Delta4 and delta4, 6-17alpha-acyloxy-20-oxo-steroid-[3, 2-c] pyrazoles of the pregnane series



United States Patent 3,148,183 A AND A -17a-ACYLOXY-Zli-OXO-S'I'EROID-[3,Z-C] PYRAZQLES 0F TEE PREGNANE SERIES Ralph F. Hirschrnann, Scotch Plains, Arthur A. Patchett, Metuchen, Avery Rosegay, Springfield, and Gerald J. Kent, Princeton, N..l., assignors to Merck & (30., Inc., Rahway, N.J., a corporation of New Jersey No Drawing. Filed June 20, 1962, Ser. No. 203,748 16 Claims. (Cl. 260-2395) This invention is concerned generally with novel [3,2-c1pyrazole compounds of the pregnane series, and with processes of preparing the same. More particularly, it relates to 17a-hydroxy-ZO-oXo-4-pregneno- (and 4,6- pregnadieno)-[3,2-c]pyrazoles, and to processes of making these compounds. It also relates to the 17u-ethers and 17a-acyloxy-derivatives of the 17a-hydroxy-4-pregnone (and 4,6-pregnadieuo) [3,2-c] pyrazoles.

These novel [3,2-c1pyrazole compounds may be chemiwherein R is selected from the group consisting of hydrogen and methyl, R is selected from the group consisting of hydrogen, a-methyl, a-fluoro and m-ChIOIO, R is selected from the group consisting of hydrogen, u-fluo-ro, a-methyl, B-methyl and methylene, R is selected from the group consisting of hydrogen, hydroxy, acyloxy and fiuoro, R is selected from the group consisting of hydrogen, alkyl, cycloalkyl, aralkyl, aryl, a heterocyclic nucleus or substituted derivatives thereof, and R is selected from the group consisting of hydrogen, acyl, alkyl, aralkyl and alkenyl. This invention also includes the A -analogues of all of the above compounds.

The above defined [3,Z-cJpyr-azole compounds are intermediates in the production of the corresponding 11(3- hydr0xy-[3,2-c]pyrazoles by microbiological oxidation. The 1IB-hydroxy-[3,2-c]pyrazoles possess high anti-inflammatory activity and are especially elfective for the treatment of arthritis and related diseases since they can be administered for their cortisone-like action in low dosage, thereby minimizing undesirable side effects.

ice

The 17a-hydroxy-20-oxo-4-(pregneno and 4,6-pregnadieno)-[3,2-c]pyrazoles are prepared by the method described in Flow Sheet A, or by the method described in Flow Sheet B. For obtaining a particular compound, the choice of method may be determined by the availability of the starting materials, among other considerations. The starting materials for our invention can also be prepared by the introduction of the various substituents, namely the 16a-methyl-, 16fi-methyl-, 16-methylene-, 6a-methyl-, 6oc-ChlO1O-, and 6oc-fl110r0- into the 17a-hydroXy-4-pregnene-3,20-dione or 17a-hydroxy 4,6- pregnadiene-3,20-dione (Flow Sheet A), or the 1704,21- dihydroxy-4-pregnene-3,2O dione or 1711,21 dihydroxy- 4,6-pregnadiene-3,20-dione (Flow Sheet B), using known procedures capable of general application.

Following the synthesis described in Flow Sheet A, the 17a-hydroXy-4-pregnene-(or 4,6-pregnadiene)-3,20-dione is converted into the 17ot-tetrahydropyranyl ether. This may be accomplished by refluxing under anhydrous conditions with dihydropyran in the presence of an acidic reagent such as |tosyl chloride.

The l7a-tetrahydropyranyloxy 4 pregnene (or 4,6- pregnadiene)-3,20-dione (Compound 2) is reduced to the corresponding 17u-tetrahydropyranyloxy-4-pregnene (or 4,6-pregnadiene)-3,20-diol, for example, by heating with lithium aluminum hydride in a solvent such as tetrahydrotur'an. The l7a-tetrahydropyranyloxy 4 pregnene- (or 4,6-pregnadiene)-3,20-diol (Compound 3), is then oxidized to 1700 .tetrahydropyranyloxy 20 hydroxy 4- pregnene-(or 4,6-pregnadiene) -3 one, conveniently by the use of manganese dioxide.

Upon treatment of the 17a-tetrahydropyranyloxy-20- hydroxy-4-pregnene-(or 4,6-pregnadiene) 3 one (Compound 4) with an alkyl formate and sodium hydride in an inert atmosphere there is formed the corresponding 2- hydroxymethylene-derivative. The Z-hydroxymethylene derivative (Compound 5) reacts with hydrazine in an inert atmosphere to form the corresponding 4-pregneno- (or 4,6-pregnadieno)-[3,2-c]pyrazole (Compound 6).

Upon treatment of the 2-hydroxymethylene-4-pregnene-(or 4,6-pregnadieno)-3-one (Compound 5) with a lower alkanol in the presence of an acidic reagent, such as a p-toluenesulfonic acid, the corresponding 2-alkoxymethylene-derivative is formed. When the latter compound is reacted with a monosubstituted hydrazine, the corresponding N-substituted-4-pregneno-(or 4,6 pregnadieno)-[3,2-c]pyrazole compounds (Compounds A and B) are formed. The N-substituted-4-pregneno-(or 4,6- pregnadieno)-[3,2-c1pyrazoles having structure A are designated as the 1-substituted-4-pregneno-[3,2- c]pyrazoles, and the N-substituted-pyrazole compounds having structure B are designated as the 2-substituted-4-pregneno- [3 ,2-c] pyrazoles.

wherein R R R and R have the significance above defined, and R is tetrahydropyranyl. The corresponding 4,6-pregnadieno-[3,2-c1pyrazoles are similarly designated as the l'- and 2' substituted 4,6 pregnadieno-[3,2-c] pyrazoles.

Upon treatment of a Z-hydroxymethylene-compound directly with a monosubstituted arylhydrazine, without the intermediate formation of the 2-alkoxymethylene compound, one isomer is generally formed in preponderant amounts, Whereas when reacting the 2-alkoxymethylene compound with a monosubstituted arylhydrazine, significant amounts of both isomers are obtained. When these reactions take place with mono-substituted alkylhydrazines, mixtures may be obtained starting with the Z-hydroxymethylene-steroid as well as with the Z-alkoxymethylene-steroid. A mixture of isomers may also result from the reaction of a monosubstituted hydrazine with a 2-hydroxymethylene-compound which possibly contains variable amounts of the 2-alkoxymethylene-compound due to the operating procedures employed, for example, due to recrystallization in the presence of a trace of alcohol a solution of the 2-hydroxymethylene-compound from which has not been completely removed.

Among the monosubstituted hydrazines which may be used for the process of our invention are: alkylhydrazines, such as methylhydrazine, ethylhydrazine, propylhydrazines, butylhydrazines, [i-hydroxyethylhydrazine, cycloalkylhydrazines; arylhydr-azines including phenylhydrazine and the substituted phenylhydrazines, such as o-, m-, and p-halophenylhydrazines, o-, m-, and p-tolylhydrazines, o-, m-, and p-alkoxyphenylhydrazines, 0-, m-, and p-nitrophenylhydrazines, l-hydrazinonaphthalene, Z-hydrazinopyridine, 3-hydrazinopyridine, 4-hydrazinopyridine, 4-hydrazinopyridine oxide, Z-hydrazinopyrimidine 2- and 3-hydrazinothiophene; aralkylhydrazines, such as benzylhydrazine and phenylethylenehydrazine.

There are thus produced the corresponding [3,2-c1pyrazoles including: N-alkyl such as N-methyl-, N-ethyl-, N- butyl-, N-propyl-, N-(fi-hydroxyethyD-g N-cycloalkyL', N- arylwhich may be derived from any aromatic nucleus, including N-phenyland the N-substituted-phenyl derivatives such as 0-, m-, and p-halophenyl; o-, m-, and p-tolyl-; o-, m-, and p-alkoxyphenyl-, o-, m-, and p-nitrophenyl-; N-(1"-naphthyl)-, N-(2-pyridyl)-, N-(3"-pyridyl)-, N- (4-pyridyl)-, N-(4"-pyridyloxide)-, N-(2-pyrimidyl)-, N-(2-thiophene)- and N-(3"-thiophene)- N-aralkyl-, such as N-benzyland N-phenylethenyl-pyrazole compounds. The N-alkyl-pyrazole compounds may also be prepared by direct alkylation of the N-unsubstitutedpyrazole compounds.

The 17a tetrahydropyranyloxy 20 hydroxy-4-pregneno-(or 4,6-pregnadieno)-[3,2-c]pyrazoles (Compound VI-A or VI-B of Flow Sheet A) are oxidized to the corresponding l7a-tetrahydropyranyloxy-20-oxo-4-pregneno- (or 4,6-pregnadieno)-[3,2-c]pyrazoles, suitably by using a chromium trioxide-sulfuric acid reagent which is prepared by dissolving chromium lrioxide in concentrated sulfuric acid, and then diluting With water.

The 17a-tetrahydropyranyloxy-20-oxo-4-pregneno- (or 4,6-pregnadieno)-[3,2-c1pyrazole (Compound 7) is converted into the corresponding l7a-hydroxy-derivative (Compound 8) by dissolving in methanol and treating with a solution of p-toluenesulfonic acid monohydrate in methanol.

Following the synthesis described in Flow Sheet B, a l7a,2l-dihydroxyl-pregnene (or 4,6-pregnadiene)- 3,20- dione starting material (Compound 1) is reacted with aqueous formaldehyde solutions in the presence of strong acid to form compounds having the formula:

and the A -analogues of the above compounds, wherein R is methyl or hydrogen, R is selected from the group consisting of hydrogen, a-methyl-a-chloro and a-fiuoro, and R is selected from the group consisting of a-rnethyl, (i-methyl, methylene and u-fluoro. For example, cold concentrated HCl and formalin are added to a stirred suspension of the steroid in chloroform, cooled to about 0 C. The mixture is then allowed to come to room temperature and stirred for several hours to afford the corresponding 20, 20, 21-bis(methylenedioxy)-derivative.

Upon treatment of the above compound With an alkyl formate and sodium hydride in an inert atmosphere there is formed the corresponding 17a, 20, 20, 2l-bis(methylenedioxy) 2-hydroxymethylene-4-pregnene-(or 4,6-pregnadiene)-3-one (Compound 3). In a preferred embodiment of our invention the steroid is dissolved in a solvent such as benzene and the resulting solution is cooled to room temperature and treated with ethyl formate. The air in the system is replaced with nitrogen, sodium hydride is added and the mixture is stirred at room temperature for several hours.

The 170;, 20, 20, 21-bis(methylenedioxy)-2-hydroxymethylenel-pregnene (or 4,6-pregnadiene)-3-one compound reacts With hydrazine in an inert atmosphere to form the corresponding 170:, 20, 20, 21-bismethylenedioxy) 4- pregneno (or 4,6 pregnadieno) [3,2 c]- pyrazole.

Upon treatment of a 170:, 20, 20, 21-bis(methylenedioxy) 2 hydroxymethylene 4 pregnene (or 4,6- pregnadiene)-3-one compound with a lower alkanol in the presence of an acidic reagent such as a p-toluenesulfonic acid the corresponding 17a, 20, 20, 2l-bis- (methylenedioxy) 2 alkoxymethylene 4 pregnene- (or 4,6-pregnadiene)-3-one is formed. When the latter compound is reacted with a monosubstituted hydrazine, the corresponding N-substituted-4-pregneno-(or 4,6- pregnadieno)-[3,2-c1pyrazoles (Compounds 4A and 4B) are formed. The N-substituted-[3,2-c]pyrazoles having structure A are designated as the l'-substituted-[3,2-c]- pyrazoles, and the N-substituted-pyrazole compounds having structure B are designated as the 2-substituted- [3,2-61-pyrazoles; in these formulas the linkage between carbons 6 and 7 may be a single or double bond linkage.

wherein R R and R have the significance above defined, and R is hydrogen, alkyl, cycloalkyl, aryl, aralkyl, a heterocyclic nucleus, or substituted derivatives thereof. The products formed may be separated by chromatography.

Upon treatment of a 17a, 20, 20, 2l-bis(methylenedioxy)-2-hydroxymethylene compound directly with a monosubstituted arylhydrazine, without the intermediate formation of the 2-alkoxymethylene-derivative, one isomer is generally formed in preponderant amounts, whereas when reacting the 170:, 20, 20, 21-bis(methylenedioxy)-2-alkoxymethylene-4-pregnene-3 -one compound with a monosubstituted arylhydrazine, significant amounts of both isomers are obtained. When these reactions take place with monosubstituted alkylhydrazines, mixtures may be obtained when starting with the Z-hydroxymethylenesteroid as well as with the 2-alkoxymethylene-steroid. A mixture of isomers may also result from the reaction of a monosubstituted hydrazine with a 2-hydroxymethylene-compound which possibly contains variable amounts of the 2-alkoxymethylene-derivative due to the operating procedures employed, for example, due to recrystallization in the presence of a trace of alcohol a solution of the Z-hydroxymethylene-compound from which acid has not been completely removed.

The monosubstituted hydrazines which may be used for the process of Flow Sheet B are those listed in column 3.

Upon treatment of any of the above described 170:, 20, 20, 21-bis(methylenedioxy)-[3,2-c]pyrazole compounds (Compounds 4A and 4B) with a dilute organic acid, for example, a 60% aqueous solution of formic acid, the 171:, 20, 20, 21-bis(methylenedioxy)-protecting group is removed and there is obtained the corresponding 17oz, ZI-dihydroxy-ZO-oxo[3,2-c]pyrazoles which are represented by Compounds 5A and 5B.-

onion;

Rs-N

wherein the linkage between carbons 6 and 7 may be a single or double bond linkage and wherein R is hydrogen or methyl; R R and R have the meaning described above, and R is hydrogen or an acyl group corresponding to the organic acid used in this reaction. An acyl group may also be present at R when the reactant (Compound 4) is an unsubstituted pyrazole.

Any acyl groups present at R; and/or at R may be removed by treating the steroid with sodium methoxide in methanol at room temperature. Acyl groups present at the R position may be selectively removed by treatment with aqueous acetic acid. Acylating agents which can be used for this purpose include a lower hydrocarbon carboxylic acid acylating agent such as benzoic anhydride, tertiary butyl acetyl chloride; a lower alkanoic anhydride or lower alkanoyl halide such as acetic anhydride, propionic anhydride or acetyl chloride; or a polybasic anhydride such as 18,/3-dimethyl-glutaric anhydride, succinic anhydride and the like, in the presence of an organic base such as pyridine.

The 17a, 2l-dihydroxy-[3,2-c1pyrazole is reacted with methane sulfonyl chloride in a non-aqueous base to form the 21-mesylate (Compound 6). A steroid in which R is hydrogen is preferably converted to the N-carbamyl derivative before undergoing this reaction.

The 2l-fiuoro-l7a-hydroxy-pyrazoles (Compound 8) are prepared from the corresponding 17u,21-dihydroxy- [3,2-c1pyrazole 2l-mesylates by heating with an alkali fluoride in a solvent to form a mixture of a 1711,21- expoxy-compound and the corresponding 21-fluoro-compound. These compounds are separated by partition chromatography, or by chromatography on a weak adsorbent such as silica gel.

The 1706, 21-dihydroxy-[3,2-c]pyrazole 2l-mesylate is heated with an alkali iodide to form the 21-iodo-compound (Compound 7). In a preferred method for carrying out this reaction, sodium iodide is added to the steroid dissolved in acetone and the resulting mixture is heated at reflux temperature for approximately one hour.

The l7ot-hydroxy-21-iodo-[3,2-c]pyrazole is heated with an alkali bisulfite in a solvent to form the corresponding 21-desoxy-compound (Compound 9). A preferred method is to add sodium bisulfite to a suspension of the steroid in aqueous ethanol and then heat the mixture under reflux for a period of about an hour.

An N-carbamyl group is removed by treatment with nitrous acid. For example, the steroid is treated in glacial acetic acid solution with sodium nitrite.

All of the [3,2-c]pyrazoles described in the foregoing structures form salts such as the hydrochloride, sulfate, chlorate, perchlorate, picrate and trichloroacetate, on treatment with the corresponding acid. Formation of crystalline salts, especially the hydrochloride salts, is sometimes helpful as a means of purifying the [3,2-c1pyrazoles.

Acylation of the above pyrazole products with pyridine and an acyl anhydn'de leads to 2l-acyl derivates except when R is hydrogen, in which case N,2l-diacyl derivatives result. Optionally, the latter derivatives may be funther heated with dilute aqueous acetic acid to remove selectively the N-acyl group.

The l'Ia-acyl-derivatives of the l7u-hydroxy-4-pregneno- (and 4,6-pregnadieno)-[3,2-c1pyrazoles are prepared by acylation of the corresponding l7a-hydroxycompound.

The 17a-hydroxy-compound is readily acylated by reaction with a lower aliphatic acid anhydride at elevated temperatures. In general, it has been found necessary to carry out this l7a-acylation step at temperatures in excess of about 100 C. for extended periods of time. Thus, the acylation can most conveniently be achieved by refluxing the reaction mixture for a period in excess of about 5 hours, and generally for about -12. hours for optimum results.

Alternatively instead of employing high temperatures to carry out this step, acylation of the 17ot-hydroxycompound may be achieved at room temperature by reaction with an acid anhydride in the presence of an acid catalyst such as mineral acids, for example, hydrochloric acid, sulfuric acid and the like, or a sulfonic acid such as p-ttoluene suifonic acid. With the latter catalyst, the reaction time at room temperature is generally about eighteen hours.

As acylating agents it is desirable to use such aliphatic acid anhydrides as acetic anhydride, propionic acid an hydride or butyric acid anhydride, although it will be understood that similar acid anhydrides derived from other carboxylic acids may likewise be employed in this acylation procedure.

The l7a-acyloxy derivatives obtained in accordance with this process may be recovered from the reaction mixture by diluting it with water, and cooling the mixture until the desired product is precipitated. Alternatively, the reaction mixture can be concentrated in vacuo and the resulting residue triturated with Water and filtered to obtain the precipitated acyloxy compound.

When the [3,2c]pyrazole is unsubstituted, (R is hydrogen) the R hydrogen is also replaced by the corresponding acyl group in the above described acylation reaction. The acyl group at R may then be selectively removed without removal of the acyloxy group at the Not-position by heating with sodium hydroxide for about 30 minutes (See Flow Sheet C).

The flat-ethers of the l7a-hydroxy-20-oxo-4-pregneno- (and 4,6-pregnadieno)-[3,2-c]pyrazoles are prepared by contacting the l7a-hydroxy-steroid (Compound 1 of Flow Sheet A) with a monohalogenated hydrocarbon in which the halogen substituent is either bromine or iodine in the presence of a mild base such as silver oxide. Included among the halogenated hydrocarbons which may be used in the process of our invention are saturated alkyl bromides or iodides including cycloalkyl as, for example, methyl iodide, ethyl iodide, n-propyl iodide, npentyl iodide, ethyl bromide, n-propyl bromide, isopropyl bromide, n-pentyl bromide and bromocyclohexane; aralkyl bromides and iodides as, for example, benzyl iodide and benzylbromide; and unsaturated halogenated hydrocarbons such as allyl iodide, and methallyl iodide. By selecting one of the above-named halogenated hydrocarbons and reacting it with a selected 17u-hydroxy steroid of the pregnane series in accordance with the process of my invention, there is formed the corresponding l7a-ether of the starting l7a-hydroxy steroid in which the hydrogen of the 17a-hydroxy group is replaced by a hydrocarbon radical derived from the selected hydrocarbon reactant.

In order to bring about etherification of the l7m-hydroxy group it is necessary to employ, in addition to the halogenated hydrocarbon, a mild base such as silver oxide. An especially desirable form of silver oxide is prepared by the following procedure: 1 part by weight of silver nitrate is dissolved in approximately 1i) parts by weight of water. Another solution is prepared by dissolving O.23 part by weight of pure sodium hydroxide in approximately 10 parts by weight of water. The two solutions are heated to approximately 86 C. and mixed, whereupon the silver oxide precipitates. The supernatant liquid is decanted and the silver oxide precipitate is washed by decantation with 5 portions of hot water or until the washings are essentially neutral. The precipitate of silver oxide is then filtered and air-dried. The precipitated, dried silver oxide is then slurried in about 5 parts of hot ethanol. The ethanol is decanted from the precipitate and the precipitate is then washed several more times with hot ethanol by decantation. The washed precipitate is filtered and air-dried, and then dried in a desiccator over phosphorus pentoxide.

The solvent used for carrying out the etherification reaction may be any one which is inert or unreactive under the conditions of the etherification reaction. Thus, solvents which are effective as diluents are hydrocarbons as, for example, benzene, toluene, xylene; ethers such as diethyl ether, dioxane, tetrahydrofuran and anisole; N,N- dialkyl acylamides as, for example, N,N-dimethylacetamide, N,N-dimethylformamide and N,N-diethylformamide, and nitriles such as acetonitrile. In addition to solvents which may act as diluents for the reagent, the monohalogenated hydrocarbon may act as a solvent for the reaction and, when it is desired to use large excesses of the monohalogenated hydrocarbon reactant, it may be desirable to use, instead of an additional solvent, the reagent itself as a solvent for the reaction.

. The temperature at which the reaction is carried out is partially dependent on the time and does not appear to be the sole critical factor in the yield or purity of the product obtained. Thus, temperatures which vary from approximately 5 C. to the reflux temperature of the solvent employed appear to be satisfactory; Thus, we have found that when the reactants are mixed in the presence of a solvent at approximately 25 C. for a period of about 16 hours, good yields of product are obtained. Another method which has been found to produce satisfactory yields of the ether involves refluxing the steroid in the presence of silver oxide and methyl iodide for approximately 7 hours with no additional solvent added to the reaction mixture.

In a preferred mode of operation the 17rx-methoxy steroid is prepared from the corresponding 17u-hydroxy steroid by mixing the steroid with a large molar excess of methyl iodide and dimethylforrnamide as a solvent in the presence of silver oxide. The mixture is stirred at room temperature for about 4 days. The product is recovered by extraction and chromatography.

The l7a-methoxy-ether of the 4-pregnene-3,20-dione is specifically named for purposes of discussion to illustrate the process of our invention, although it is to be understood that the process is likewise applicable to all of the l7oc-BlhGf-COI1'1PO11HdS defined by Compound 2 of Flow Sheet A. The l7oc-methoxy-4-pregnene-3,ZO-dione (2) is reduced to the corresponding 17a-methoXy-4-pregnene- 3,20-diol (3), for example by heating with lithium aluminum hydride in a solvent such as tetrahydrofuran. The

17a-methoxy-4-pregnene-3,20-diol (3), is then oxidized to 17amethoxy-20-hydroxy-4-pregnene3-one (4), for example, with the use of manganese dioxide.

Upon treatment of the 17u-methoxy-20-hydroxy-4-pregnene-3-one (4) with an alkyl formate and sodium hydride in an inert atmosphere there is formed the corresponding 2-hydroxymethylene-derivative The 2-hydroxymethylene compound (5) reacts with hydrazine in an inert atmosphere to form the corresponding 4-pregneno-[3,2-c]- pyrazoles (6).

Upon treatment of the 2-hydroxymethylene-4-pregnene- 3-one compound with a lower alkanol in the presence of an acidic reagent such as a p-toluenesulfonic acid the corresponding 2 alkoxymethylene 4-pregnene-3-one is formed. When the latter compound is reacted with a monosubstituted hydrazine, the corresponding N-substituted [3,2-c1pyrazole compounds are formed (6A and 6B). The N-substituted-[3,2-c1pyrazoles having structure A are designated as the 1'-substituted-[3,2-c1pyrazoles, and the N-substituted-pyrazole compounds having structure B are designated as the 2-substituted-[3,2-c]pyrazoles.

R1 N N I wherein R R R and R have the significance above defined, and R stands for alkyl, alkenyl or aralkyl. The products formed may be separated by chromatography. The corresponding A -pregnadiene are similarly designated as the 1'- and 2'-substituted-4,6-pregnadiene-[3,2-c]- pyrazoles.

Upon treatment of a 2-hydroxymethylene-compound directly with a monosubstituted arylhydrazine, without the intermediate formation of the Z-alkoxymethylene-derivative, one isomer is generally formed in preponderant amounts, whereas when reacting the 2-a1koxymethylene with a monosubstituted arylhydrazine, significant amounts of both isomers are obtained. When these reactions take place with monosubstituted alkylhydrazines, mixtures may be obtained when starting with the Z-hydroxymethylone-steroid. A mixture of isomers may also result from the reaction of a monosubstituted hydrazine with a 2- hydroxymethylene-compound which possibly contains variable amounts of the 2-alkoxymethylene-derivative due to the operating procedures employed, for example, due to recrystallization in the presence of a trace of alcohol a solution of the Z-hydroxymethylene-compound from which acid has not been completely removed.

Among the monosubstituted hydrazines which may be used for the process of our invention are those listed on page 6.

The novel [3,2-c]pyrazoles of the present invention are converted to the corresponding 11 8-hydroxy-[3,2-c] pyrazole by the microbiological oxidation procedure of Example 28.

FLOW SHEET B 5A and 5B 6A and 6B I 7A and 7B R i I 10A and 10B I 8A and 8B 9A and 9B FLOW SHEET C Compound 8 of Flow Sheet A 10 Compounds 8 and 9 of Flow Sheet B 11 ll 3 FLOW SHEET CContinued rRr (ll zRl 0 0 Compounds A and B of Page 1 In the formulas in the preceeding Flow Sheets A, B and C the linkage between carbons 6 and 7 may be a single or double bond linkage; R is selected from the group consisting of hydrogen and methyl; R is selected from the group consisting of hydrogen, tat-methyl, a-fiuoro and a-ChlOIO; R is selected from the group consisting of hydrogen, a-methyl, ,B-methyl, methylene, and OL-fluOIO; R is selected from the group consisting of hydrogen, hydroxy, acyloxy and fiuoro; R is selected from the group consisting of hydrogen, alkyl, cycloalkyl, aralkyl, aryl and a heterocyclio nucleus, or substituted derivatives thereof; and R is selected from the group consisting of hydrogen, alkyl, aralkyl, alkenyl and tetrahydropyranyloxy.

The following examples illustrate methods of carrying out the present invention but it is to be understood that these examples are given for purposes of illustration and not of limitation.

Example 1 A solution of g. of 17a-hydroxy-4-pregnene-3,20- dione in 3 g. of tosyl chloride and 150 ml. of dihydropyran, B.P. 86 C., distilled from sodium hydride, is refluxed for 18 hours. After cooling, 100 ml. of 10% sodium carbonate solution is added, and the mixture is stirred for 1 hour. The organic layer is separated, washed with saturated salt solution, dried over magnesiurn sulfate, and concentrated under reduced pressure to a viscous residue. The residue is dissolved in 50 ml. of a 4:1 mixture of n-hexane-benzene and chromatographed on basic alumina. The product is crystallized by dissolving in 2 parts of methylene chloride and adding about 16 parts of n-hexane. Successive processing of the mother liquor yields a total of 7.1 g. of 17atetrahydropyranyloxy-4-pregnene-3,20-dione, M.P. about l55-65 C.

Example 2 A mixture of 3.6 g. of l7a-tetrahydropyranyloxy-4- preg'nene-3,2"0-dione, 2.0 g. of lithium aluminum hydride and 100 ml. of tetrahydrofuran (distilled over lithium aluminum hydride) is stirred under reflux for 1 hour. About 40 ml. of solvent is then distilled, and the mixture is cooled and treated with ethyl acetate to destroy the excess sodium hydride. With continued stirring, saturated salt solution is added in small portions until a granular deposit of salt is formed. The mixture is filtered, and the filtrate is concentrated under reduced pressure to give a 17a-tetrahydropyranyloxy-4-pregnene- 3',20-diol, which is a colorless, non-crystalline residue and shows no carbonyl absorption in the infrared.

Example 3 The 17a-tetrahydropyranyloxy-4-pregnene 3 320 diol obtained in Example 2 is dissolved in 40 ml. of acetone, and 30 g. of activated manganese dioxide is added. The mixture is stirred for 18 hours at room temperature, filtered through infusorial earth (Celite), and the manganese dioxide is washed thoroughly with methylene chloride. The combined filtrates are concentrated under reduced pressure. The residue is dissolved in ether and 14 chromatographed on basic alumina. The column is washed with ether until the eluate is free of residue, and then the product is eluted with a 1:9 solution of chloroform-ether. Evaporation of the solvent leaves a 66% yield of a colorless glass, 17a-tetrahydropyranyloxy- 20-hydroxy-4-pregnene-3-one,

Example 4 Ethyl formate 1.5 ml.) is added to a solution of 1.63 g. of 17oc-tetrahydropyranyloxy-lfl-hydroxy-4-pregnene- 3-one in 35 ml. of dry benzene, and the system is flushed with dry nitrogen. Sodium hydride (1.5 g. as a 53% oil dispersion) is added, and after flushing again with nitrogen, the mixture is stirred for 2 hours at room temperature. After cooling, 30 ml. of ether and 30 ml. of a saturated solution of sodium dihydrogen phosphate solution are added, and the mixture is shaken for about 5 minutes. The layers are separated, the water layer is washed with ether, and the combined ether extracts are Washed with saturated salt solution. The product is then extracted with six 15 ml. portions of 2% sodium hydroxide solution and the combined basic extracts are washed three times with ether. The alkaline solution is acidified to a pH of 6 using saturated sodium dihydrogen phosphate solution, and then extracted several times with ether. After washing the combined ether extracts with saturated salt solution and then drying over sodium sulfate, partial evaporation of solvent gives 400 mg. of crude, crystalline product, M.P. 152-5 C. The mother liquor is evaporated to dryness and the residue is dissolved in 2 ml. of ether to afford a second crop (450 mg.) of product, M.P. 153-5 C. The total yield of crude product is about 49% )G (in base) 357 me, E% 231; 242 mu, E% 344 Several recrystallizations from methylene chloride-ether gives an analytical sample of 20-hydroxy-2-hydroxymeth ylene-17a-tetrahydropyranyloxy-4-pregnene-3 one, M.P. 185-9 C.,

AESQ (in base) 357 my, 13% 240; 243 mu, E% 363 Example 5 A solution of 3.5 g. of 17a-tetrahydropyranyIOXy-Z0- hydrox-2-hydroxymethylenet-pregnene-3-one and 0.45 ml. of hydrazine hydrate in 30 ml. of ethanol is refluxed under nitrogen for 45 minutes. The solvent is then evaporated under reduced pressure. The residue is flushed with n-hexane and ml. of ether is added. The resulting suspension is shaken thoroughly for several minutes, filtered, and the filtrate evaporated to dryness under reduced pressure, to leave a residue of about 1.6 g. (46%) of 17a-tetrahydropyranyloxy-20-hydroxy- 4-pregneno-[3,2-c1pyrazole,

260 mu, E% 201, 9,000

The ether insoluble fraction shows max.

max.

Example 6 A mixture of 1 gram of l7a-tetrahydropyranyloxy-2 hydroxy-Z-hydroxymethylene-4-pregnene-3one, 200 ml. of methanol, and 200 mg. of p-toluensulfonic acid is heated to reflux temperature and then allowed to stand at room temperature for one hour. The reaction mixture is then diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed two times with 2 N aqueous sodium hydroxide solution and then with Water. The ethyl acetate extract is then dried and concentrated in vacuo. The 17a-tetrahydropyranyloxy-ZO- hydroxy-Z-methoxymethylene-4 pregnene 3 one is obtained by chromatography on acid-washed alumina and elution with ether:chloroform mixtures.

A mixture of -00 mg. of the above Z-methoxymethylone-derivative, 100 ml. of ethanol, and 1 ml. of methylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature overnight. Acetic acid (2 ml.) is added and the mixture is allowed to stand for another 4 hours. The reaction mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extract is then dried, concentrated, and chromatographed on acid-washed alumina to yield the 1'-methyland the 2'-methyl-17a-tetrahydropyranlyoxy-ZO-hydroxy-4-pregneno-[3,2-c1pyrazole.

The N methyl 17a tetrahydropyranyloxy 20'- hydroxy-4-pregneno-[3,2-c]pyrazole may also be prepared by the following procedure: A solution of about 0.47 millimole of 17wtetrahydrOpyranyIoXy-ZO-hydroxy- 4-pregneno-[3,2-c]pyrazole, in ml. of benzene is treated with 30-38 mg. of about 51% sodium hydride (in oil suspension) after the addition of 23 ml. of dimethyl formamide (dried over calcium hydride) and 5 ml. of methyl iodide, the mixture is stirred at room temperature over night. The product is filtered, washed with methylene chloride, and the filtrate and washing are taken to dryness. The residue is treated with water and the product is filtered to give N-methyl-17a-tetrahydropyranyloxy--hydroxy-4-pregneno- [3 ,2-c] pyrazole.

In accordance with the above procedures, but using other alkylating agents in place of methyl iodide, there is obtained the corresponding N alkyl 171x tetrahydropyranyloxy-Z 0-hydroxy-4-pregneno- [3 ,2-c] pyrazole.

In accordance with all of the above procedures, but starting with the Z-hydroxymethylene derivatives defined by Compound 5 of Flow Sheet A, there are obtained the corresponding l-methyland 2'-methyl derivatives.

Example 7 A mixture of 90 mg. of 17u-tetrahydropyranloxy-ZO- hydroxy-Z-hydroxymethylene-4-pregnene-3-one and 0.028 ml. of phenylhydrazine is refluxed under nitrogen in 1.2 m1. of absolute ethanol for about 50 minutes. The reaction mixture is taken to dryness. Water is added and the product is filtered to give an amorphous solid, which is washed successively with water, dilute acid, water, and petroleum ether. The product is crystallized from methanol to afford a product which is predominately the 2 phenyl 17 a tetrahydropyranyloxy 20 hydroxy- 4-pregneno- [3 ,=2-c] pyrazole.

A mixture of the 1'-phenyland 2'-phenyl-17a-tetrahydropyranyloxy 20 hydroxy 4 pregneno [3,2 -c]- pyrazole is prepared by the following route: A mixture of 1 gram of 17a-tetrahydropyranyloxy-20-hydroxy-2-hydroxymethylene-4-pregnene-3-one, 200 ml. of methanol, and 200 mg. of p-toluenesulfonic acid is heated to rcfiux temperature and then allowed to stand at room temperature for one hour. The reaction mixture is then diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed two times with 2 N aqueous sodium hydroxide solution and then with water. The ethyl acetate extract is then dried and concentrated in vacuo. The 170; tetrahydropyranyloxy 20 hydroxy-2- methoxymethylene-4-pregnene-3-one is obtained by chromatography on acid-washed alumina and elution with etherzchloroform mixtures.

A mixture of 509 mg. of the above Z-methoxymethylenesteroid, ml. of ethanol, and 1 ml. of phenylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature over night. Acetic acid (2 ml.) is added and the mixture is allowed to stand for another 4 hours. The reaction mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extract is then dried, concentrated, and chromatographed on acid-washed alumina to yield the l'-phenyland 2'-phenyl-l7a-tetrahydropyranyloxy-ZO- hydroxy-4-pregneno- [3 ,2-c] pyrazole.

In accordance with the above procedures, but starting with the Z-hydroxymethylene-derivative which is obtained from each of the starting materials which are defined by Compound 5 of Flow Sheet A, there are obtained the corresponding l'-phenyland 2'-phenyl-derivatives.

In accordance with the above procedures, but using cyclohexylhydrazine, p-tolylhydrazine, p-chlorophenylhydrazine, p-methoxyphenylhydrazine, or benzylhydrazine in place of phenylhydrazine, there are obtained the corresponding land 2-cyclohexyl-, 1'- and 2-p-tolyl-, 1-, and 2-p-chlorophenyl-, 1'- and 2I-p-methoxyphenyl-, and 1'- and 2'-benzyl-17a-tetrahydropyranyloxy-20-hydroxy- 4-pregneno-[3,2-c]pyrazoles.

Example 8 A 111.5 mg. sample of l7a-tetrahydropyrany1oxy-20- hydroxy-2-hydroxymethylene-4-pregnene-3-one is suspended in 2.5 ml. of ethanol and treated with 24.5 mg. of sodium acetate, followed with the addition of 48.5 mg. of p-fluorophenylhydrazine hydrochloride. The air in the system is replaced with nitrogen and the mixture is quickly brought to reflux temperature. After refluxing for one hour the mixture is taken to dryness. The residue is dissolved in ether, the ether layer is treated three times with 2.5 N hydrochloric acid. then three times with 2.5 N sodium hydroxide and finally with water. The ether layer is dried over magnesium sulfate, filtered and concentrated to dryness in vacuo. The product is dissolved in methanol and then allowed to crystallize slowly to afford a product which is predominantly the 2 (p fluorophenyl) 17oz tetrahydropyranyloxy 20- hydroxy-4-pregneno- [3 ,-2-c] pyrazole.

A mixture of 1'-(p-fluorophenyl)- and 2'-(p-fluorophenyl) 17 a tetrahydropyranyloxy 2 01 oxo 4 pregneno-[3,2-c]pyrazole is prepared by the following route: A mixture of 1 gram of l7a-tetrahydropyranyloxy-20- hydroxy-Z-hydroxymethylene-4-pregnene-3-one, 200 ml. of methanol, and 200 mg. of p-toluenesulfonic acid is heated to reflux temperature for one hour. The reaction mixture is then diluted with water and extracted with ethyl acetate. The ethyl acetate extract is Washed two times with 2 N aqueous sodium hydroxide solution and then with water. The ethyl acetate extract is then dried and concentrated in vacuo. The l7ec-tetrahydropyranyloxy 20 hydroxy 2 methoxymethylene 4 pregnene- 3-one is obtained by chromatography on acid-washed alumina and elution with ether-chloroform mixtures.

A mixture of 500 mg. of the 2-methoxymethylenederivative 100 ml. of ethanol, and 1 ml. of p-fluorophenylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature over night. Acetic acid (2 ml.) is added and the mixture is allowed to stand for another 4 hours. The reaction mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extracts are then dried, concentrated, and chromatographed on acid-washed alumina to afford the 1-(p fluorophenyl) and 2'-(p-fiuorophenyl)17a-tetrahydropyranyloxy-ZO-hydroxy-4-pregneno- 3,2-c1pyrazole.

In accordance with all of the above procedures, but starting with the 2-hydroxymethylene derivative which is obtained from each of the starting materials which are defined by Compound 5 of Flow Sheet A, there are obtained the corresponding 1'-(p-fluorophenyl)- and 2'- (p-fluorophenyl)-derivatives.

Example 9 The 17a-tetrahydropyranyloxy-ZO-hydroxy-4-pregneno- [3,2-c1pyrazole (326.0 mg.) is dissolved in 32.6 ml. of purified acetone and cooled in an ice bath to 05 C. A chromium trioxide-sulfuric acid reagent (0.247 ml.), prepared by dissolving 26.72 g. of chromium trioxide in 23 cc. of concentrated sulfuric acid and then diluting With water to a volume of 100 cc., is added and the mixture is stirred for 40 seconds. Cold 0.1 N sodium hydroxide (20 ml.) is added, followed by 170 m1. of cold water. The mixture is stirred at room temperature for 10 minutes. The product is filtered, washed with water and air-dried for 10 minutes. It is then dissolved in 40 ml. of methanol and filtered. The solvent is removed in vacuo. The residue is then dissolved in 10 ml. of chloroform and the solution is dried over magnesium sulfate, filtered and the solvent removed on a steam bath. On drying 264.7 mg. of residue is obtained. The residue is chromatographed on silica gel. The fractions which have been eluted with 99% chloroform-1% methanol are collected and combined with chloroform. The solvent is removed to give 108.3 mg. of 17a-tetrahydropyranyloxy- 20-oxo-4-pregneno-[3,2-c]pyrazole A 260 11111., E% 177. The LR. spectrum shows some 17 carbonyl to be present.

Example 10 The 17a-tetrahydropyranyloxy 20 oxo 4 pregneno- [3,2-c]pyrazole (40.0 mg.) is dissolved in 0.5 ml. of methanol and treated with 1.82 ml. of a solution of 500 mg. of p-toluenesulfonic acid monohydrate in 25 ml. of methanol. The mixture is kept at room temperature for 4 hours. The solvent is removed on a steam bath and the residue is treated with 3 ml. of ethyl acetate. The insoluble material is filtered off and the mother liquor washed twice with 2 m1. of 10% sodium bicarbonate and twice with 2 m1. of water. The product is dried over magnesium sulfate, filtered and the solvent removed on a steam bath. The residue is slurried with a small amount of methylene chloride, and the slurry is filtered and dried in vacuo to give 23.0 mg. of 17a-hydroxy- 20-oxo-4-pregneno-[3,2-c1pyrazole U.V. X1 52 260 my, E% 293 In accordance with the above procedure, but starting with an N-substituted-l7u-tetrahydropyranyloxy-ZO-oxo- 4-pregneno'[3,2-c1pyrazole the corresponding N-substituted-17a-hydroxy-20-oxo-4-pregneno- [3 ,2-c] pyrazole is obtained.

Example 11 The 17cc hydroxy 20-oxo-4-pregneno-[3,2-c1pyrazole (37.3 mg.) is treated with 3 ml. of acetic anhydride and 1 ml. of a solution of 304 mg. of p-toluenesulfonic acid monohydrate dissolved in 10 ml. of acetic anhydride. The mixture is kept at room temperature for 20 hours. The solvent is removed in vacuo. The residue is dissolved in 1 ml. of ether which has been washed once with 1 ml. of water. The aqueous phase is back-extracted twice with 1 ml. of ether. The combined ethereal extracts are washed once with 2 ml. of 10% sodium bicarbonate solution. The aqueous phase is then backextracted with 1 ml. of ether. The combined ether extracts are washed with 1 ml. of water. The aqueous extract is back extracted with 1 ml. of ether. The combined ethereal extracts are dried over magnesium sulfate, filtered and the solvent removed on a steam bath to give 18 42.2 mg. of crude 17a-acetoxy-20-oxo-4-pregneno-[3,2-c] pyrazole N-acetate.

Example 12 The 17a-acetoXy-20-oxo-4-pregneno-[3,2-c1pyrazole N- acetate is dissolved in 3.0 ml. of methanol and treated with 0.13 ml. of 0.1 N sodium hydroxide. The mixture is heated at reflux temperature for 30 minutes. The solvent is removed on a steam bath to give a crude product which is treated with 1 ml. of Water and then extracted with 1 ml. of ether. The layers are separated and the aqueous phase is back extracted three times with 1 ml. of ether. The combined ether extracts are washed with 1 ml. of water. The organic phase is dried over magnesium sulfate, filtered and the solvent removed. The product is crystallized from methanol to give 23.0 mg. of 17a-acetoxy-ZO-oxo-4-pregneno- 3,2-c] pyrazole $52 260.0 mg., E% 236 (9,400) LR. CHClgOAC and NH of pyrazole ring 2.89 3.1, 3.2a.

Following in sequence the procedures of Examples 1, 2, 3, 4, 5, 9, 10, 11 and 12, but starting in Example 1 with each of the following compounds in place of the 17u-hydroxy-4-pregnene-3,20-dione; 17a-hydroxy-19-nor-4-preg-nene,3,20 dione, 17a-hydroxy-6,16a-dirnethyl-4,6-pregnadiene-3,20-dione, 17u-hydroxy-4,6-pregnadiene-3,20-dione, 17 u-hydroxy-6-chloro1 6a-methyl-4,6-pregnadiene- 3,20-dione, 17a-hydroxy-6,16fl-dimethyl-4,G-pregnadiene-3,20-dione, 17 a-hydroxy-6-ch1oro-4,6-pregnadiene-3,20-dione,

there is obtained as a product, the corresponding [3,2-01- pyrazole, namely: i

17 a-acetoxy- 19-nor-20-oxo-4-pregneno- 3,2-c] pyrazole,

17a-acetoXy-6,16a-dimethyI-ZO-oxo-4,6-pregnadieno- [3,2-c] pyrazole,

17u-acetoxy-20-oxo-4,6-pregnadieno- [3,2-c] -pyrazole,

17a-acetoxy-6-chloro-16a-methyl-20-oxo-4, 6-pregnadieno- [3,2-c] pyrazole,

17a-acetoxy-6,16,8-dimethyl-20-oxo-4,6-pregnadieno- [3,2-c] pyrazole,

17a-acetoxy-6-chloro-20-oxo-4,6-pregnadieno- 3,2-c]

pyrazole.

Following in sequence the procedures of Examples 1, 2, 3, 4, 7, 9, 10 and 11, but starting in Example 1 with each of the following compounds in place of the 17a-hydroxy-4-pregnene-3,20 dione; 17a-hydroxy-6a-methyl-4-pregnene-3,20-dione, 17u-hydroxy-6-chloro-1 6a-methyl-4,6-pregnadiene-3,20-

dione, 17ot-hydroxy-6,16a-dimethyl-4,6-pregnadiene-3,20-dione, 17a-hydroxy-6-l6fl-dimethyl-4,6-pregnadiene-3,20-dione, 17a-hydroXy-6-methyl-4,G-pregnadiene-3,20-dione, 17 u-hydroxy-6-chloro-4,6-pregnadiene-3,20-dione,

there is obtained as a product the corresponding 2'-phenyl- [3,2-c] pyrazole namely:

l7a-acet0xy-6a-methyl-20-oxo-2'-phenyl-4-pregneno- [3,2-c] pyrazole, 17u-acetoxy-6-chloro-16a-methyl-20-oxo-2'-phenyl-4,6-

pregnadieno- [3,2-c] pyrazole, 17a-acetoxy-6,16a-dimethyI-ZO-oxo-2'-phenyl-4,6-

pregnadieno- 3,2-c] pyrazole, 17u-acetoxy-6,16fl-dimethyl-20-oxo-2'-phenyl-4,6- pregnadieno- 3,2-c] -pyrazole, 17u-acetoxy-6-methyl-20-oxo-2-phenyl-4,6-pregnadieno- [3,2-c] pyrazole, 17a-acetoxy-6-chloro-20-oxo-2-phenyl-4,6-pregnadieno- [3,2-c]pyrazo1e.

The 17a-hydroxy-6-chloro-16a-methyl-4,6-pregnadiene- 3,20-dione used as starting material can be made by the following procedures:

A solution of 3.25 g. of 17a-hydroxy-16a-methyl-4,6- pregnadiene-3,20-dione in 325 ml. of methylene dichloride is cooled in an ice bath and treated with 75 ml. of 1.5 N ethereal monoperphthalic acid and allowed to stand over night at room temperature. The mixture is poured into excess sodium bicarbonate, and the organic layer is separated and dried. Removal of solvent and crystallization from a suitable solvent affords 6a,7a-epoxy-17a-hydroxy-l 6u-methyl-4-pregnene-3,20-dione.

6a,7a-epoxy-17a-hydroxy-16a-methyl-4-pregnene-3,20- dione (1.0 g.) in 75 ml. of glacial aceticacid is sat urated at room temperature with anhydrous hydrogen chloride and allowed to stand for four hours at room temperature. It is then poured into water, extracted with chloroform, washed with water and sodium bicarbonate. Removal of dried solvent and chromatography on neutral alumina affords 6-chloro-17a-hydr0xy-16a-methyl-4,6 pregnadiene-3,20-di one.

Example 13 A solution of 1.000 g. of 17a-hydroxy-20-oxo-4-pregneno-[3,2-c]pyrazole in 10 ml. of caproic acid and 2 ml. of trifluoroacetic acid is flushed with argon and heated on a steam bath for 50 minutes. The mixture is then cooled, poured into water and the acid is neutralized with sodium bicarbonate. The product is extracted with methylene chloride. The extract is washed with 5% sodium hydroxide solution and water, dried and concentrated to an oil. The oil is chromatographed on Florisil (an activated magnesium silicate made according to U.S.P. 2,393,625) to obtain the 17a-caproyloxy-20-oxo-4-pregneno-[3,2-c]- pyrazole.

Following in sequence the procedures of Examples 1, 2, 3, 4, 5, 9, and 13, but starting with 17u-hydroxy- 6,16tx-dimethyl-4,6-pregnadiene-3,ZO-clione there is obtained the l7a-caproyloxy-6,16a-dirnethyl-20 oxo 4,6- pregnadieno-[3,2-c]pyrazole.

Example 14 To a suspension of 25.0 g. of 17a,21-dihydroxy-16amethyl-4-p regnene-3,20-dione in 1.5 l. of alcohol-free chloroform cooled to about 5 C. in an ice bath is added with constant stirring 750 ml. of cold, concentrated hydrochloric acid and then 750 ml. of formalin (low in methanol). The mixture is removed from the ice bath and stirred at room temperature for 7 hours. The layers are separated and the aqueous phase is back-extracted twice with chloroform. The combined organic layers are washed twice with a 5% solution of sodium bicarbonate, and twice with a saturated salt solution. The solution is dried over magnesium sulfate and evaporated to dryness under reduced pressure. The residue is triturated with methanol to aflord a crystalline solid. This material contains no detectable amount of starting material by paperstrip chromatography but shows two U.V. absorbing spots near the solvent front (methanol-fonnamide 2:1 vs. benzene-n-hexane 1:1). A 2.425 g. aliquot is recrystallized three times from a mixture of benzene and n-hexene to give 17a,20,20,2l-bis(methylenedioxy)-16 x-methyl-4pregnene-3-one which is used in the subsequent step of the synthesis without further purification.

The 17u,20,20,21 bis(methylenedioxy) 16a methyl- 4-pregnene-3-one (1.350 g.) is dissolved in 23 ml. of dry, hot benzene and the resulting solution is cooled to room temperature and treated with 0.96 ml. of freshly distilled ethyl formate. The air in the system is replaced with nitrogen and 560 mg. of sodium hydride (as a 58% dispersion in mineral oil) is added. The system is again evacuated and filled with nitrogen, and the mixture is stirred magnetically at room temperature over night. The mixture is poured into an excess of a saturated aqueous solution of sodium dihydrogen phosphate and the product is extracted four times with benzene. The organic extracts are washed three times with water and dried over sodium sulfate. Removal of the solvent gives the crude product which is dissolved in ether and purified as the sodium salt by extraction into a 10% solution of sodium carbo nate. The aqueous alkaline extracts are again acidified with an excess of a saturated aqueous solution of sodium dihydrogen phosphate and extracted into ether and into chloroform. The combined organic extracts are dried over sodium sulfate and evaporated to dryness to give 17a,20,20,2l bis(methylenedioxy) 2 hydroxymethylene-16ot-methyli-pregnene3-one.

The l7a,Z0,20,21 bis(methylenedioxy) 2 hydroxymethylene-16a-methyl-4-pregnene-3-one (850 mg.) is dis solved in 9.2 ml. of absolute ethanol and treated with a solution of 0.16 ml. of hydrazine hydrate dissolved in 0.16 ml. of absolute ethanol. The mixture is refluxed in a nitrogen atmosphere for about 45 minutes and then evaporated to dryness under reduced pressure. The residue is washed three times with cold water and the resulting amorphous solid is dried at C. for 1 hour in high vacuum to give l7a,20,20,2l-bis(methylenedioxy)-l6 xmethyl-4-pregneno- 3,2-c] pyrazole.

The l7u,20,20,21 bis(methylenedioxy) 16a methyl- 4-pregneno-[3,2-c1pyrazole (720 mg.) is heated in a steam bath with 24 ml. of a 60% aqueous solution of formic acid for about 30 minutes. The excess reagent is removed in vacuo using a water bath at about 50 C. as the source of heat. The residue is flushed four times with n-hexane and then dried at 60 C. in high vacuum to give an amorphous solid which is a mixture of 170;,21- dihydroxy 16cc methyl 20 oxo 4 pregneno-[3,2-c]- pyrazole and/or its formylated derivatives. A 500 mg. aliquot of this crude product is dissolved in 2.4 ml. of pure methanol and allowed to react with 0.9 ml. of a 1.33 N solution of sodium methoxide in methanol at room temperature under a nitrogen atmosphere for ten minutes. The alkoxide is neutralized with acetic acid and the mixture is then taken to dryness and flushed with n-hexane. The residue is washed with water, filtered and dried to constant weight to give 17e,2l-dihydroxy-l6cx-methyl-20- oxo-4-pregneno-[3,2-c1pyrazole.

To a solution of mg. of l7u,21-dihydroxy-16amethyl-20-oxo-4-pregneno-[3,2-c]pyrazole in 2 ml. of pyridine is added two milliequivalents of acetic anhydride. The mixture is allowed to stand overnight at room temperature. A mixture of ice and water is then added. After standing for about 30 minutes, the product is extracted with ethyl acetate. The ethyl acetate extract is washed successively with water, ice-cold 1 N sulfuric acid (until the pH of the aqueous layer is l3), saturated aqueous sodium bicarbonate (until the pH of the aqueous layer is 8), and water (until the aqueous layer is neutral). The ethyl acetate solution is dried with anhydrous sodium sulfate. The solvent is then distilled at about 40 C. in vacuo to aiford the N-acetyl-17a,2l-dihydroxy 16a methyl 20 oxo 4 pregneno-[3,2-c]- pyrazole 2l-acetate which is isolated by the addition of Water and filtration.

In accordance with the above procedure, but using an equivalent quantity of another acylating agent in place of acetic anhydride, there is obtained the corresponding N acyl 17,21 dihydroxy 16a methyl 20 oxo 4- pregneno- 3,2-c] pyrazole 21-acylate.

A solution of 5.73 g. of N-acetyl-l7or,21-dihydroxy-l6umethyl-20oxo-4-pregneno-[3,2-c]pyrazole 21-acetate in 60 ml. of 80% (v./v.) acetic acid is refluxed for 1.5 hours. This solution is diluted with 400 ml. of ice-water and extracted with ethyl acetate. The ethyl acetate extracts are washed with water and with saturated sodium bicarbonate solution, dried, and evaporated to dryness in vacuo. Recrystallization of the resulting product atfords the l7a-21-dihydroxy-l6o-methyl-20 oxo 4 pregneno- [3,2-c1pyrazole Zl-acetate.

To a solution of 3.70 g. of 17a,21-dihydroxy-16amethyl-20-oxo-4-pregneno-[3,2-c]pyrazole in 150 ml. of methanol there is added 30 ml. of water containing 0.01 mole of hydrogen chloride. Then a solution of 0.81 g. of potassium cyanate in 8 ml. of water is added and this mixture is left over night at room temperature. Some of the methanol is removed in vacuo on the rotating evaporator; more water is added and the precipitate is collected by filtration. Recrystallization from methanol gives the N-carbamyl pyrazole in sufficient purity for the next step.

To a solution of 85 m of N-carbamyl-lhQl-dihydroxy 16m methyl 20 oxo 4 pregneno-[3,2-c]pyr azole in 0.5 ml. of pyridine, cooled to C., is added 0.015 ml. of methane sulfonyl chloride. The resulting mixture is allowed to stand at a temperature of approximately 0 C. for a period of approximately one hour. Water is then added to the reaction mixture and the precipitate which forms is recovered by filtration, washed with water, and dried to give N-carbamyl-17,21-dihydroxy- 16a-methyl-20-oxo-4-pregneno- [3 ,2-c] pyrazole 21- mesylate.

To a solution of 62 mg. of N-carbamyl-17a,2l-dihydroxy 16 methyl 20 oxo 4 pregneno-[3,2-c]pyrazole 21-mesylate in 1 ml. of freshly distilled anhydrous dimethylformamide is added enough anhydrous potassium fluoride to assure a saturated solution. The mixture is heated at 110 C. for 20 hours. Water is added to the cooled solution and the product is extracted into chloroform, dried over sodium sulfate and evaporated to dryness. The resulting product is a mixture of N-carbamyl- 170:,21 epoxy 16oz methyl 20 oxo 4 pregneno- [3,2-c1pyrazole and N-carbamyl-Z1-fiuoro-17ct-hydrox 16cc methyl 20 oxo 4 pregneno [3,2 c]pyrazole, which compounds are separated by partition chromatograph or by chromatography on silica gel.

To a solution of 355 mg. of N-carbamyl-Zl-fiuoro-Uahydroxy 16a methyl-20-oxo-4-pregnen0-[3,2-0]pyrazole in 35 ml. of glacial acetic acid there is added slowly with stirring 104 mg. of sodium nitrite in 5 ml. of water. After fifteen minutes at room temperature, most of the acetic acid is removed at room temperature on the rotating evaporator. Ethyl acetate is added and this solution is extracted several times with sodium bicarbonate and then dried. Removal of the solvent, followed by chromatography on alumina, aifords 2l-fluoro-17a-hydroxy-16otmethyl-ZO-oxo-4-pregneno- 3,2-c] pyrazole.

To 180 mg. of N-carbamyl-17a, 21-dihydroxy-16umethyl-20-oxo-4-pregneno[3,2-c] pyrazole 2l-mesylate dissolved in ml. of acetone is added 300 mg. of sodium iodide. The resulting mixture is heated at reflux temperature for a period of approximately 1 hour, and the reaction solution is cooled to room temperature and diluted with water. The material which precipitates is recovered by filtration, washed with water, and dried to give N- carbamyl l7ix-hydroxy 21 iodo-16u-methyl-20-oxo-4 pregneno-[3,2-c]pyrazole.

The N carbamyl-l7a-hydroxy-21-iodo-16u-methyl-20 oxo-4-pregneno-[3,2-c1pyrazole is dissolved in a mixture of 5 ml. of water and 5 ml. of ethanol. To the resulting suspension is added 500 mg. of sodium bisulfite and the mixture is heated under reflux for a period of about 1 hour. The reaction solution is cooled, diluted with water, and the material which separates is recovered by filtration. The product is washed with water, dried and recrystallized from ethyl acetate to give N-carbamyl- 17a hydroxy-16u-methyl-20-oxo-4-pregneno-[3,2-c1pyrazole.

To a solution of 355 mg. of N-carbamyl-lM-hydroxy- 20-oxo-4-pregneno-[3,2-c]pyrazole in 35 ml. of glacial acetic acid there is added slowly with stirring 104 mg. of sodium nitrite in 5 ml. of water. After fifteen minutes at room temperature, most of the acetic acid is removed at room temperature on the rotating evaporator. Ethyl acetate is added and this solution is extracted several times 22 with sodium bicarbonate and then dried. Removal of the solvent, followed by chromatography on alumina, alfords 17a-hydroxy-20-oxo 4-pregneno-[3,2-c]pyrazole.

Example 15 To a solution of 0.5 millimole of 17a, 20, 20, 21-bis (methylenedioxy) 2 hydroxymethylene-16a-methyl-4- pregnene-3-one in about 3 ml. of absolute ethanol is added 0.6 millimole of sodium acetate and then 0.6 millimole of methylhydrazine sulfate. The mixture is refluxed under nitrogen for 40 minutes and then filtered hot. The filtrate is taken to dryness, water is added, and the 1704, 20, 20, 2l-bis (methylenedioxy-N, 16u-dimethyl-4-pregneno-[3,2-c]pyrazoie which is formed as the major component is removed by filtration.

Alternately, a mixture of 1', 16a-dimethyl-, and 2, 16adimethyl-17a, 20,20,21-bis(methylenedioxy)-4-pregneno- [3,2c]pyrazole is prepared by heating 17oz, 20,20,21-bis (methylenedioxy) 2 hydroxymethylene 16a-methyl-4- pregnene-3-one with methanol in the presence of ptoluenesulfonic acid to form the 17a,20,20,21-bis(methylenedioxy -2-methoxymethylene-16a-methyl-4-pregnene-3 one, and then reacting the latter compound with methylhydrazine, following the detailed procedures given in column 2.4, but using methylhydrazine instead of phenylhydrazine in the second step of the reaction. The components of the mixture are separated by chromatography.

In accordance with all the above procedures, but using other alkyl substituted hydrazines such as ethyl-, fi-hydroxyethyl-, propyl-, butylhydrazines, and the like, in place of methylhydrazine, there are obtained the corresponding 1'-alkyland 2-alkyl-17ot,20,20,21-bis(methylenedioxy) -l6a-methyl-4-pregneno- [3 ,2-c1pyrazoles.

The 17u,2U,Z0,21-bi$ (methylenedioxy)-N, 16ot-dimethyl enedioxy) N, 16ot-dimethyl-4-pregneno-[3,2-c]pyrazoles may be prepared also by the following procedure:

The l7oc,20,20,21 bis(methylenedioxy) 2 hydroxymethylene-16ot-n1ethyl-4-pregnene-3-one is dissolved in 9.2 ml. of absolute ethanol and treated with a solution of 0.16 ml. of hydrazine hydrate dissolved in 0.16 ml. of absolute ethanol. The mixture is refluxed in a nitrogen atmosphere for about 45 minutes and then evaporated to dryness under reduced pressure. The residue is washed three times with cold water and the resulting amorphous solid is dried at C. for one hour in high vacuum to give 17a,20,20,21 bis (methylenedioxy) 16oz methyl-4- pregneno- 3 ,2-c] pyrazole.

A solution of about 0.47 millimole of 17a,20,20,21-bis (methylenedioxy) 16a methyl-4-pregneno-[3,2 c]pyrazole in 10 m1. of benzene is treated with about 30-38 mg. of about 51% sodium hydride (in oil suspension). After the addition of 2-3 ml. of dimethylformamide (dried over calcium hydride) and 5 ml. of methyl iodide, the mixture is stirred at room temperature overnight. The product is filtered, washed with methylene chloride, and the filtrate and washings are taken to dryness. The residue is treated with Water and the product is filtered to afford as a major component the 17a,20,20,21 bis(methylenedioxy)-N, 16cc dimethyl-4-pregneno- [3 ,2-c] pyrazole.

In accordance with the above procedure, but using another alkylating agent, for example, ethyl iodide, propyl iodide and the like, in place of the methyl iodide, there is obtained the corresponding N-alkyl-17a,20,20,21-bis- (methylenedioxy)-16a-methy1-4-pregneno [3,2-c]pyrazole.

The 17a,20,20,21-bis(methylenedioxy)-2,16oc-dimethyl-4-pregneno-[3,:2-c]pyrazole (15 mg.) is heated on a steam bath with 1 ml. of 60% formic acid for about 20 minutes and then filtered hot. The filtrate is taken to dryness, water is added, and a mixture of the 17a,21-dihydroxy-2',16a dimethyl-ZO-oxo-4-pregneno-[3,2-c1pyrazole and its 2l-formate is recovered by filtration. A 500 mg. aliquot of this crude product is dissolved in 2.4 ml. of pure methanol and allowed to react with 0.9 ml. of a 1.33.N solution of sodium methoxide in methanol at room temperature under a nitrogen atmosphere for ten minutes. The alkoxide is neutralized with acetic acid and the mixture is then taken to dryness and flushed with nhexane. The residue is washed with Water, filtered and dried to constant weight to give 17a,2l-dihyd10Xy-2',l6ocdimethyl-20-oxo4-pregneno- [3,2-c1pyrazole.

' To a solution of 100 mg. of l7u,2l-dihydroxy-2',l6adimethyl 20 oxo-4-pregneno-[3,2-c1pyrazole in 2 ml. of pyridine is added one milliequivalent of acetic anhydride. The mixture is allowed to stand overnight at room temperature. A mixture of ice and Water is then added. After standing for about 30 minutes, the prodnot is extracted with ethyl acetate. The ethyl acetate extract is washed successively with water, ice-cold l N sulfuric acid (until the pH of the aqueous layer is 1-3), saturated aqueous sodium bicarbonate (until the pH of the aqueous layer is 8), and water (until the aqueous layer is neutral). The ethyl acetate solution is dried with anhydrous sodium sulfate. The solvent is then distilled at about 40 C. in vacuo. The l7a,2l-dihydroxy-2'16u-dimethyl-20-oxo-4-pregneno [3,2c]pyrazole Zl-acetate is then isolated by addition of water and filtration.

In accordance with the above procedure, but using an equivalent quantity of another acylating agent in place of acetic anhydride, there is obtained the corresponding 17a,2l-dihydroxy2',l6a dimethyl-20-oxo-4 pregneno- [3,2-c1pyrazole 21-acylate.

To a solution of 85 mg. of l7a,-dihydroxy-2,l6a-dimethyl-20-oxo-4-pregneno-[3,2-c1pyrazole in 0.5 ml. of pyridine, cooled to C., is added 0.015 ml. of methane sulfonyl chloride. The resulting mixture is allowed to stand at a temperature of aproxinrately 0 C. for a period of approximately 1 hour. Water is then added to the reaction mixture and the precipitate which forms is recovered by filtration, Washed with water, and dried to give 17a,2l dihydroxy-ZZlM-dimethyl-20-oxo-4-preg- 'nen0[3,2-c]pyrazole 2l-mesylate.

To 180 mg. of l7a,21-dihydroxy-2'-lSa-dimethyl-ZO- oxo-4-pregneno-[3,2-c1pyrazole 21-mesylate dissolved in 10 ml. of acetone is added 300- mg. of sodium iodide. The resulting mixture is heated at reflux temperature for a period of approximately 1 hour, and the reaction solution is cooled to room temperature and diluted with water. The material which precipitates is recovered by filtration, washed with water, and dried to give 17 a-hydroxy- 21-iodo-2,l6a-dirnethyLZO-oxol-pregneno [3,2-c1pyrazole.

The 170: hydroxy-2l-iodo-2',16a-dinethyl-20-oxo-4- pregneno-[3,2-c]pyrazole is dissolved in a mixture of ml. of water and 5 ml. of ethanol. To the resulting suspension is added 500 mg. of sodium bisulfite, and the mixture is heated under reflux for a period of about 1 hour. The reaction solution is cooled, diluted with water, and the material which separates is recovered by filtration. The product is Washed with water, dried and recrystallized fr-orn ethyl acetate to give 17a-hydroxy-2,16adimethyl 20 oxo-4-pregnenp-[3,2-c1pyrazole.

To a solution of 62 mg. of 17a,21-dihydroxy-2',16adimethyl-20-oxo-4-pregneno-[3,2-c]pyrazole 21-mesylate in 1 m1. of freshly distilled anhydrous dimethyl-formamide is added enough anhydrous potassium fluoride to assure a saturated solution. The mixture is heated at 110 C. for 20 hours. Water is added to the cooled solution and the product is extracted into chloroform, dried over sodium sulfate and evaporated to dryness. The resulting product is a mixture of l7a,2l-epoxy-2',l6a-dimethyl-20-oxo-4- pregneno [3,2-c1pyrazole and 21-fluoro-17a-hydroxy- 2,l6a-dimethyl 20 oxo-4-pregneno-[3,2-c]pyrazole, which compounds are separated by partition chromatography, or by chromatography on silica gel.

In accordance with the above procedures beginning with the 2'-methyl-17a,20,20,2l-bis(methylenedioxy)-4- pregneno-[3,2-c]pyrazole, but using the l-methy1-derivative in place of the 2'-methyl-derivative, there are obtained the corresponding 1'-methyl compounds.

24 In accordance with all of the above procedures, but starting with the 17a,20,20,21-bis(methylenedioxy)-2-hydroxymethylenet-pregnene-(or 4,6-pregnadiene) which is obtained from each of the starting materials defined by Compound 3 of Flow Sheet B, there are obtained the corresponding 1-rnethyland 2-methyl derivative.

Example 16 A mixture of mg. of 17a,20,20,2l-bis(methylenedioxy)-2-hydroxymethylene l6a methyl-4-pregneno-3- one and 0.028 ml. of phenylhydrazine are refluxed under nitrogen in 1.2 ml. of absolute ethanol for 50 minutes. The reaction mixture is taken to dryness. Water is added and the product is filtered to give an amorphous solid, which is washed successively with water, dilute acid, water, and petroleum ether. The product is crystallized from methanol to give l7zx,20,20,2l-bis(methylenedioxy)- l6a-methyl-2'-phenyl-4-pregneno-[3,2-c1pyrazo1e.

Alternately, a mixture of the l'-phenyland 2'-phenyl- 17 11,20,202 l-bis(rnethylenedioxy)- l 6a-methyl-4-pregneno- [3,2-c1pyrazole is prepared by the following route: A mixture of 1 gram of l7oz,20,20,2l-bis(methylenedioxy)- 2-hydroxymethylene-l6a-methyl-4-pregnene-3-one, 200 ml. of methanol, and 200 mg. of p-toluenesulfonic acid is heated to reflux temperature and then allowed to stand at room temperature for one hour. The reaction mixture is then diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed two times with 2 N aqueous sodium hydroxide solution and then with water. The ethyl acetate extract is then dried and concentrated in vacuo. The l7a,20,20,2l-bis(methylenedioxy)- Z-methoxymethylene-l6a-methyl-4-pregnene-3-one is obtained by direct crystallization or by chromatography on acid-Washed alumina and elution with etherzchloroform mixtures.

A mixture of 500 mg. of l7ct,20,20,2l-bis(methylenedioxy)-2-methoxymethylene 16a methyl 4 pregnene- 3-one, m1. of ethanol, and 1 ml. of phenylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature overnight. Acetic acid (2 ml.) is added and the mixture is allowed to stand for another 4 hours. The reaction mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extract is then dried, concentrated, and chromatographed on acidwashed alumina. Elution with benzene atlords a product which on recrystallization from a mixture of benzene and ethyl acetate alfords the l7a,20 .20,2l-bis(methylenedioxy) 16cc methyl l phenyl-4-pregneno-{3,2-c]pyrazole. Further elution with a mixture of 1:1 ether: petroleum ether and crystallization from benzene, afiords the l7e,20,20,21-bis(rnethylenedioxy)-l6oa-methyl-2'-phenyl-4-pregneno-[3,2-cjpyrazole.

A 30 mg. aliquot of l7a,20,20,2l-bis(methylenedioxy)- 16e-methyl-2-phenyl-4-pregneno-[3,2-c1pyrazole is heated on a steam bath with 2 ml. of 60% formic acid for 35 minutes. The solvents are removed in vacuo, water is added and the product is filtered off to give a mixture of l7a,21-dihydroxy-l6oz-methyl-20-oxo-2'-phenyl-4-pregneno-[3,2-c]pyrazole and its Zl-formate. The presence of formate is indicated by infrared absorption at 5.81 and 8.5a.

A 500 mg. aliquot of this crude product is dissolved in 2.4 ml. of pure methanol and allowed to react with 0.9 ml. of a 1.33 N solution of sodium methoxide in methanol at room temperature under a nitrogen atmosphere for 10 minutes. The alkoxide is neutralized with acetic acid and the mixture is then taken to dryness and flushed with n-hexane. The residue is washed with water, filtered and dried to constant weight to give 1701,21- dihydroxy-l6a-methyl-20 oxo 2' phenyl 4 pregneno- [3,2-c1pyrazole.

The l7a,2l-dihydroxy-l6a-methyl-20-oxo-2-phenyl-4- pregneno-[3,2-c]pyrazole is treated with a mixture of 1.5 ml. of pyridine and 1.5 ml. of acetic anhydride and the mixture is allowed to stand at room temperature over night. The solvents are removed in vacuo, water is added and the 17a,21-dihydroxy-l6a-methyl-20-oxo-2'- phenyl-4-pregneno-[3,2-c1pyrazole 21 acetate is removed by filtration. After drying, the compound is dissolved in methylene chloride, a few drops of 2.5 N HCl are added and the mixture is taken to dryness. The resulting 17a,2l-dihydroxy-16a-methyl-ZO-oxo-2'-phenyl-4-pregneno-[3,2-c]pyrazole Zl-acetate hydrochloride salt is soluble in methylene chloride and can be crystallized from acetone.

The 17a,2l-dihydroxy-l6ot-methyl-20-oxo-2-phenyl-4- pregneno-[3,2-c]pyrazole (100 mg.) is dissolved in 1.2 cc. of dimethylformamide. The solution is cooled to C. and 0.07 cc. of methane sulfonyl chloride is added. The mixture is kept at 0 C. for about one hour, 3 cc. of water is added and the product is extracted into 100 cc. of chloroform, Washed with water, dried over sodium compounds.

In accordance with all of the above procedures, but starting with the 17a,20,20,21-bis(methylenedioxy)-2- hydroxymethylene-4-pregnene derivative which is obtained from each of the starting materials defined by Compound 3 of Flow Sheet B, there are obtained the corresponding 1'-phenyland 2'-phenylderivatives.

Example 17 A 111.5 mg. sample of 17a,20,20,21-bis(methylenedioxy) 2 hydroxymethylene l6a-methyl-4-pregnene-3- one is suspended in 2.5 ml. of ethanol and treated with 24.5 mg. of sodium acetate, followed with the addition of 48.5 mg. of p-fluorophenylhydrazine hydrochloride. The air in the system is replaced with nitrogen and the mixture is quickly brought to reflux temperature. After refluxing for one hour the mixture is taken to dryness. The residue is dissolved in ether, the ether layer is treated three times with 2.5 N hydrochloric acid, then three times with 2.5 N sodium hydroxide and finally with water. 'The ether layer is dried over magnesium sulfate, filtered and concentrated to dryness in vacuo to give a residue which has as its major component the l7oc,20,20-2l-bi$- (methylenedioxy) 2 (p fluorophenyl) l6a-methyl-4- pregneno-[3,2-c]pyrazole. The latter compound is recovered by dissolving the reaction mixture in methanol and then recrystallizing.

Alternately, a mixture of the l'-(p-fluorophenyl)- and the 2-(p-fluorophenyl)-17a,20,20,2l-bis(methylenedioxy)- l6a-methyl-4-pregneno-[3,2-c1pyrazole is prepared by the following route: A mixture of 1 gram of 17tx,20,20,21- bis(methylenedioxy) -2-hydroxymethylene 16a methyl- 4-pregnene-3-one, 200 ml. of methanol, and 200 mg. of p-toluenesulfonic acid is heated to reflux temperature and then allowed to stand at room temperature for one hour. The reaction mixture is then diluted with Water and extracted with ethyl acetate. The ethyl acetate extract is Washed two times with 2 N aqueous sodium hydroxide solution and then with water. The ethyl acetate extract is then dried and concentrated in vacuo. The 17m,20,20,2l-bis(methylenedioxy) 2 methoxymethylene-16a-methyl-4-pregnene-3-one is obtained by direct crystallization or by chromatography on acid-Washed alumina and elution with etherzchloroform mixtures.

A mixture of 500 mg. of 17a,20,20,21-bis(methylenedioxy) 2 methoxymethylene 16a methyl 4 pregnene-3-one, 100 m1. of ethanol, and 1 ml. of p-fluorophenylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature overnight. Acetic acid (2 ml.) is added and the mixture is allowed to stand for another 4 hours. The reaction mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extract is then dried, concentrated, and chromatographed on acid-washed alumina.

Elution with benzene affords a product which on recrystallization from a mixture of benzene and ethyl acetate afiords the 17a,20,20,21 bis(methylenedioxy)- 16u-methyl-1'-fluorophenyl 4 pregneno[3,2-c] pyrazole. Further elution with a mixture of 1:1 etherspetroleum ether, and crystallization from benzene, aifords the l7a,20,20,21 bis(methylenedioxy) 16a methyl-2'- (p-fluorophenyl -4-pregneno- 3,2-c] pyrazole.

A 70 mg. aliquot of 17u,20,20,21-bis(methylenedioxy) 2'-(p-tluorophenyl) 16a methyl-4pregneno-[3,2-c] pyrazole is heated on a steam bath with 2 ml. of a 60% solution of formic acid for 35 minutes. The excess reagent is removed in vacuo using a water bath at about 50 C. The residue is thoroughly washed with water and then dried at C. to give 61.1 mg. of residue. The crude product is dissolved in 3 ml. of spectral grade methanol and allowed to react with 0.5 ml. of a 0.1 N solution of sodium methoxide in methanol at room temperature for 10 minutes. The product is neutralized with acetic acid. The mixture is then taken to dryness and washed thoroughly with water, filtered and dried to constant weight to give l7ot,21-dihydroxy-2'-(p-fluorophenyl)-16a-methyl-ZO-oxo-4-pregneno [3,2-c] pyrazole.

To a solution of mg. of 17a,21-dihydroxy-2-(pfluorophenyl) 16cc methyl-ZO-oxo-pregneno [3,2-c] pyrazole in 2 ml. of pyridine is added one milliequivalent of acetic anhydride. The mixture is allowed to stand over night at room temperature. A mixture of ice and water is then added. After standing for about 30 minutes, the product is extracted with ethyl acetate. The ethyl acetate is washed successively with water, ice-cold 1 N sulfuric acid (until the pH of the aqueous layer is 1-3), saturated aqueous sodium bicarbonate (until the pH of the aqueous layer is 8), and water (until the aqueous layer is neutral). The ethyl acetate solution is dried with anhydrous sodium sulfate. Thesolvent is then distilled at about 40 C. in vacuo to afford 17a,21-dihydroxy-2- (p-fiuorophenyl)-16a-methyl-ZO-oxo-4-pregneno [3,2-c] pyrazole 21-acetate which is isolated by the addition of water and filtration. After drying, the compound is dissolved in methylene chloride, a few drops of 2.5 N HCl are added and the mixture is taken to dryness. The resulting hydrochloride 'salt is crystallized from acetone.

In accordance with the above procedure, but using an equivalent quantity of another acylating agent in place of acetic anhydride, there is obtained the corresponding 21-acylate.

To a solution of 85 mg. of 2'-(p-fluorophenyl)-17u,21- dihydroxy-16a-methyl 20 oxo-4-pregneno-[3,2-c]pyrazole in 0.5 ml. of pyridine, cooled to 0 C., is added 0.015 ml. of methanesulfonyl chloride. The resulting mixture is allowed to stand at a temperature of approxi mately 0 C. for a period of approximately 1 hour. Water is then added to the reaction mixture and the crystalline precipitate which forms is recovered by filtration, washed with water, and dried to give 2'-(p-fluorophenyl)-17a,21-dihydroxy 16oz methyl-20-oxo-4-pregneno- [3,2-c] pyrazole 2 l-mesylate.

To ml. of 2'-(p-fiuorophenyl)-17a,21-dihydroxyl6a-methyl-20-oxo 4 pregneno [3,2-c]pyrazole 21- mesylate dissolved in 10 ml. of acetone is added 300 mg. of sodium iodide. The resulting mixture is heated at reflux temperature for a period of approximately 1 hour, and the reaction solution is cooled to room temperature and diluted with water. The crystalline material which precipitates is recovered by filtration, washed with water, and dried to give 2'-(p-fluorophenyl)-l7a-hydroxy-21- iodo-l6a-methyl 20 oxo-4-pregneno-[3,2-c]pyrazole.

The 2'-(p-fluorophenyl)-17a-hydroxy 21 iodo 16amethyl-20-oxo-4-pregneno-[3,2-c1pyrazole is dissolved in a mixture of 5 m1. of water and 5 m1. of ethanol. To the rwul-ting suspension is added 500 mg. of sodium bisulfite, and the mixture is heated under reflux for a period of about 1 hour. The reaction solution is cooled, diluted with water, and the crystalline material which 2? separates is recovered by filtration. The product is washed with Water, dried and recrystallized from ethyl acetate to give 2-(p-fiuorophenyl) 17a hydroxy-16a-methyl- 20-oxo-4-pregueno- I 3 ,2-c] pyrazole.

To a solution of 62 mg. of 2-(p-fiuorophenyl)-l7a,2ldihydroxy-l6zx-methyl 20 oxol-pregneno-[3,2-c1pyrazole 21-mesylate in 1 ml. of freshly distilled anhydrous dimethylformamide is added enough anhydrous potassium fluoride to assure a saturated solution. The mixture is heated at 110 C. for 20 hours. -Water is added to the cooled solution and the product is extracted into chloroform, dried over sodium sulfate and evaporated to dryness. The resulting product is a mixture of 2'-(p-fluorophenyl)-17a,21-epoxy 16a methyl-20-oxo-4-pregneno- [3,2-clpyrazole and 2-(p-fluorophenyl)-2l-fluoro 17cchydroxy-16a-methyl 20 oxo 4 pregneno-[3,2-c} pyrazole which compounds are separated by partition chromatography.

In accordance with the above procedures beginning with the 2'-(p-fiuorophenyl)-l7u,20,20,2l-bis(methylene dioxy)-6a-methyl-4-pregneno-[3,2-c]pyrazole, but using the l-(p-fiuorophenyl)-derivative in place of the 2'-(p fluorophenyl)derivative, there are obtained the corresponding 1-(p-luorophenyl)-compounds.

In accordance with all of the above procedures, but starting with the l7a,20,20,2l-bis(methylenedioxy)-2- hydroxymethylene-4-pregnene-derivative which is obtained from each of the starting materials defined by Compound 3 of Flow Sheet B, there are obtained the corresponding 1'- (p-fluorophenyl)- and 2-(p-fiuorophenyl)-derivatives.

Example 18 A mixture of 500 mg. of l7a-hydroxyl-pregnene-3,20- dione, 10 ml. of dimethylformamide, ml. of methyl iodide, and 1.5 gms. of silver oxide is stirred at room temperature for 3 days. An additional /2 gm. of silver oxide is added and the mixture is stirred for one day longer. One hundred ml. of chloroform is then added to the reaction mixturewhich is stirred for one hour and filtered. The filtrate is evaporated to dryness. The residual oil is chromatographed with acid-washed alumina and eluted with mixtures of ether and petroleum ether to give 17a-methoxy-4pregnene-3,20-dione.

In the manner described above but using ethyl iodide, allyl iodide, or benzyl iodide in place of methyl iodide, the product obtained is l7a-ethoxy-, 17a-allyloxyor 17abenzyloxy-4-pregnene-3-one.

Example 19 Example 20 The l7a-methoxy-4-pregnene-g3,20-diol is dissolved in 40 ml. of acetone and g. of activated manganese dioxide is added. The mixture is stirred for 18 hours at room temperature, filtered through infursorial earth .(Celite) and the manganese dioxide is washed thoroughly with methylene chloride. The combined filtrates are concentrated under reduced pressure. The residue is dissolved in ether and chromatographed on basic alumina. The column is Washed with ether until the eluate is free of residue, and then the product is eluted with chloroform-ether 1:9. Evaporation of the solvent leaves 20- hydroxy-17e-methoxy-4-pregnene-3-one.

28 Example 21 Ethyl formate (1.5 ml.) is added to a solution of 1.63 g. of ZO-hydroxy-l7a-methoxy-4-pregnene-3-one in 35 ml. of dry benzene, and the system is flushed with dry nitrogen. Sodium hydride (1.5 g. as a 53% oil dispersion) is added, and after flushing again with nitrogen, the mixture is stirred for 2 hours at room temperature. After cooling, 30 ml. of ether and 30 ml. of a saturated solution of sodium dihydrogen phosphate solution are added, and the mixture is shaken for about 5 minutes. The layers are separated, the Water layer is washed with ether, and the combined ether extracts are Washed With saturated salt solution. The product is then extracted with six 15 ml. portions of 2% sodium hydroxide solutiomand the combined basic extracts are Washed three times with ether. The alkaline solution is acidified to a pH of 6 using saturated sodium dihydrogen phosphate solution, and then extracted several times with ether. After Washing the combined ether extracts With saturated salt solution and then drying over sodium sulfate, partial evaporation of solvent gives a crude, crystalline product. The mother liquor is evaporated to dryness and the residue is dissolved in 2 ml. of ether to afford a second crop of ZO-hydroxy-Z-hydroxymethylene-l7oc-methoxy-4-pregnene-3-one.

Example 22 A solution of 3.5 g. of non-crystalline 20-hydroxy-2- hydroxymethylenel7oa-methoxy-4-pregnene-3-one and 0.45 ml. of hydrazine hydrate in 30 ml. of ethanol is refluxed under nitrogen for 45 minutes. The solvent is then evaporated under reduced pressure. The residue is flushed with n-hexane and ml. of ether is added. The resulting suspension is shaken thoroughly for several minutes, filtered, and the filtrate evaporated to dryness under reduced pressure, to leave a residue of 20-hydroxy-l7a methoxyl -pregneno- [3 ,2-c] pyr azole.

Example 23 is filtered, Washed with Water and air-dried for 10 minutes.

It is then dissolved in 40 ml. of methanol and filtered. The solvent is removed in vacuo. The residue is then dissolved in 10 ml. of chloroform and the solution is dried over magnesium sulfate, filtered and the solvent removed on steam bath. On drying, a residue is obtained. This is chromatographed on silica gel to give l7a-methoxy-20- oxo-4-pregneno-[3,2-c1pyrazole.

Example 24 To a solution of 0.5 millimole of 20-hydroxy-2-hydroxymethylene-l7a-methoxy-4-pregnene-3-one in about 3 ml. of absolute ethanol is added 0.6 millimole of sodium acetate and then 0.6 millimole of methylhydrazine sulfate. The mixture is refluxed under nitrogen for 40 minutes and then filtered hot. The filtrate is taken to dryness, Water is added and the 20-hydroxy-N-methyl-17amethoxy-4-pregneno-[3,2-c1pyrazole is removed by filtration.

A mixture of the l-methyland 2-methyl-20-hydroxyl7a-methoxy-4-pregneno-[3,2-c1pyrazole is prepared by the following route: A mixture of 1 gram of ZO-hydroxy- Z-hydroxyrnethylene-17a-methoxy-4-pregnene-3-one, 200 ml. of methanol, and 200 mg. of p-toluenesulfonicacid is heated to reflux temperature and then allowed, to stand at room temperature for one hour. The reaction mixture is then diluted with water and extracted with ethyl acetate.

The ethyl acetate extract is washed two times with 2 N aqueous sodium hydroxide solution and then with water. The ethyl acetate extract is then dried and concentrated in vacuo. The 20-hydroxy-l7u-methoxy-2-methoxymethylene-4-pregnene-3-one is obtained by chromatography on acid-washed alumina and elution with etherzchloroform mixtures.

A mixture of 500 mg. of the above 2-methoxymethylcue-derivative, 100 ml. of ethanol, and 1 ml. of methylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature over night. Acetic acid (2 ml.) is added and the mixture is allowed to stand for another 4 hours. The reaction mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extract is then dried, concentrated, and chromatographed on acid-washed alumina to ofiord the l-methyland the 2'-methy1-20-hydroxy-l7u-methoxy-4-pregneno- [3,2-c1pyrazole.

Alternately, the 20-hydroxy-N-methyl 17a-methoxy- 4-pregneno-[3,2-c]pyrazoles may be prepared by the following procedure: A solution of about 0.47 millirnole of 20-hydroxy-l7u-methoxy-4-pregneno-[3,2-c1pyrazole in m1. of benzene and about 30-38 mg. of about 51% sodium hydride (in oil suspension) after the addition of 2-3 ml. of dimethylformamide (dried over calcium hydride) and 5 ml. of methyl iodide, the mixture is stirred at room temperature over night. The product is filtered, washed with methylene chloride, and the filtrate and washings are taken to dryness. The residue is treated with water and the product is filtered to give -hydroxy-N- methyl-l7a-methoxy-4-pregneno- [3,2-c]pyrazo1e.

In accordance with the above procedures, but using other alkylating agents in place of methyl iodide, there is obtained the corresponding N-alkyl-20-hydroxy-l7a-methoxy-4-pregneno- [3 ,2-c] pyrazole.

In accordance with all of the above procedures, but starting with the 2-hydroxymethylene derivatives defined by Compound 5 of Flow Sheet A, there are obtained the corresponding 1'-methy1- and 2-methyl-derivatives.

Example A mixture of.90 mg. of 20-hydroxy-2-hydroxymethylene-l7a-methoxy-4-pregnene-3-one, and 0.028 ml. of phenylhydrazine are refluxed under nitrogen in 1.2 ml. of absolute ethanol for about 50 minutes. The reaction mixture is taken to dryness. Water is added and the product is filtered to give an amorphous solid, which is washed successively with water, dilute acid, water, and petroleum ether. The product is crystallized from methanol to afford a product which is predominantly the 20- hydroxy 17oz methoxy-2-phenyl-4-pregneno-[3,2 c]- pyrazole.

A mixture of 1'-phenyland 2'-phenyl-20-hydroxy-l7amethoxy-4-pregneno-[3,2-c]pyrazole is obtained by the following route: A mixture of 1 gram of '20-hydroxy-2- hydroxymethylene-l7a-methoxy-4-pregnene-3-one, 200 ml. of methanol, and 200 mg. of p-toluenesulfonic acid is heated to reflux temperature and then allowed to stand at room temperature for one hour. The reaction mixture is then diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed two times with 2 N aqueous sodium hydroxide solution and then with water.

The ethyl acetate extract is then dried and concentrated in vacuo. The 20-hydroxy-2-methoxymethylene-17amethoxy-4-pregnene-3-one is obtained by chromatography on acid-washed alumina and elution with ether:chloroform mixtures.

.A mixture of 500 mg. of the above 2-methoxy-methylene-derivative, 100 ml. of ethanol, and 1 ml. of phenylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature overnight. Acetic acid (2 ml.) is added and the mixture is allowed to stand for another 4 hours. The reac tion mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extract is then dried, concentrated, and chromatographed on acid-washed alumina to afford the l'-phenyland 2'-phenyl-20-hydroxy-l7a-methoxy-4-pregneno-[3,2- c] pyrazole.

In accordancewith the above procedures, but starting with the 2-hydroxymethylene-derivative which is obtained from each of the starting materials defined by Compound 5 of Flow Sheet A, there are obtained the corresponding l'-pheny1- and 2-pheny1 derivatives.

In accordance with the above procedures, but using other arylhydrazines, for example, p-nitrophenyl-, pmethoxyphenylhydrazine, 1hydrazinonaphthalene, 2-, 3-, or 4-hydrazinopyridine, 4-hydrazinopyridine oxide or 2- hydrazinopyrimidine, or the like, in place of phenylhydrazine, there it obtained the corresponding 1'-aryland 2-aryl-17u-methoxy-20-hydroxy-4-pregneno- [3,2-c] pyrazole.

Example 26 A 111.5 mg. sample of ZO-hydroxy-2-hydroxymethylene-l7u-methoxy-4-pregnene-3-one is suspended in 2.5 ml. of ethanol and treated with 24.5 mg. of sodium acetate, followed with the addition of 48.5 mg. of p-fiuorophenylhydrazine hydrochloride. The air in the system is replaced with nitrogen and the mixture is quickly brought to reflux temperature. After refluxing for one hour the mixture is taken to dryness. The residue is dissolved in ether, the ether layer is treated three times with 2.5 N hydrochloric acid, then three times With 2.5 N sodium hydroxide and finally with water. The ether layer is dried over magnesium sulfate, filtered and concentrated to dryness in vacuo. The product is dissolved in methanol and then allowed to crystallize slowly to afford a product which is predominantly the 2'-(p-fiuorophenyl)-20-hy droxy-17a-methoxy-4-pregneno-[3,2-c]pyrazole.

A mixture of l'-(p-fiuorophenyl)- and 2-(p-fluorophenyl)-20-hydroxy-17a-methoxy-4 pregneno [3,2-c]- pyrazole is prepared by the following route: A mixture of 1 gram of 20 hydroxy-Z-hydroxymethylene-l7rx-methoxy-4-pregnene-3-one, 200 ml. of methanol, and 200 mg. of p-toluenesulfonic acid is heated to reflux temperature and then allowed to stand at room temperature for one hour. The reaction mixture is then diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed two times with 2 N aqueous sodium hydroxide solution and then with water. The ethyl acetate extract is then dried and concentrated in vacuo. The 20-hydroxy- 17ot-methoxy-2-methoxymethylene-4-pregnene-3-one is obtained by chromatography on acid-washed alumina and elution with etherzchloroform mixtures.

A mixture of 500 mg. of the 2-methoxymethylenederivative, ml. of ethanol, and 1 ml. of p-fiuorophenylhydrazine is heated under nitrogen until dissolved, and then allowed to stand under nitrogen at room temperature over night. Acetic acid (2 ml.) is added and the mix" ture is allowed to stand for another four hours. The reaction mixture is then diluted with ethyl acetate, washed two times with 2 N sulfuric acid, two times with 2.5 N sodium hydroxide, and then two times with water. The ethyl acetate extracts are then dried concentrated, and chromatographed on acid-washed alumina to afford the 17a-methoxy-4-pregneno- 3,2-c] pyrazole;

In accordance with all of the above procedures, but starting with the 2-hydroxymethylene-derivative defined by Compound 5 of Flow Sheet A, there are obtained the corresponding l-(p-fiuorophenyl)- and 2-(p'-fluorophenyl)-derivatives.

Example 27 The ZO-hydroxy-l 7a-methoxy-2'-phenyl-4-pregneno- 3, 2-clpyrazole (326.0 mg.) is dissolved in 32.6 ml. of purified acetone and cooled in an ice bath to 0-5 C.

A chromium trioxide-sulfuric acid reagent (0.247 ml.) prepared by dissolving 26.72 g. of chromium trioxide in 23 cc. of concentrated sulfuric acid and then diluting with water to a volume of 100 cc. is added and the mixture is stirred for 40 seconds. Cold 0.1 N sodium hydroxide (20 ml.) is added, followed by 170 ml. of cold water. The mixture is stirred at room temperature for minutes. The product is filtered, Washed with water and air-dried for 10 minutes. It is then dissolved in 40 m1. of methanol and filtered. The solvent is removed in vacuo. The residue is then dissolved in 10 ml. cmoroform and the solution is dried over magnesium sulfate, filtered and the solvent removed on steam bath. On drying a residue is obtained which is chromatographed on silica gel to give 17a-methoxy-20-oXo2-phenyl 4 pregneno [3,2-c1- pyrazole.

In accordance With the above procedure, but starting with the ZO-hydroxy-l7u-methoxy-2'-(p-.uorophenyl)-4- pregneno-[3,2-c1pyrazole, there is obtained the 17tt-methoxy-ZO-oxo-Z' (pluorophenyl) 4 pregneno [3,2-c]- pyrazole.

Following the procedures of Examples 18, 19, 20, 21, 23, 25 and 27, but starting in Example 18 with the 170:- hydroxy 6,l6a-dimethyl-20-oXo-4,6-pregnadiene-[3,2-c]- pyrazole there is obtained the 17u-methoxy-6,16t-dimetl1- yl-20-oxo-2'-phenyl-4,6-pregnadieno- 3,2-c] pyrazole.

In accordance with all of the procedures of Examples 1 to 17, but starting in Example 1 with each of the starting materials defined by Compound '1 of Flow Sheet A and Compound 1 of Flow Sheet B, there are obtained the 17a-hydroxy-Z0-oxo-4-pregneno-[3,2-c1pyrazoles and the 17a-acyloxy-derivatives thereof, as well as the N-alkyl-, N-cycloalkyl-, N-aryl-, N-aralkyland N-heterocyclic derivatives, and the A -analogs of all of the aforesaid [3,2-c] pyrazole compounds.

There are likewise obtained the 17a,21-dihydroxy-20- oxo-4-pregneno-E3,2-clpyrazoles and the 2'1-fiuoro-17mhydroxy-20-oxol-pregnend[3,2-0] pyrazoles as well as the 17ocand the 21-acylox -derivatives thereof. The N- alkyl-, N-cycloalkyl-, N-aryl-, N-ara1kyl-, and N-heterocyclic derivatives are likewise produced as well as the A -analogs of all of the aforesaid. [3,2-c1pyrazole compounds.

In accordance with all of the procedures of Example 18 to Example 25, but starting in Example 18 with each of the starting materials defined by Compound 1 of Flow Sheet A and reacting with an alkyl, aralkyl or alkenyl halide to form Compound 2, the corresponding 17ot-alkyl-, 17OL-ElI'fl1ky1- or 17a-alkenyl-ethers of the 17a-hydroxy-20- oxo-4-pregneno-(or 4,6-pregnadieno)-[3,2-clpyrazo1es are formed, as well as the N-alky1-, N-cycloalkyl-, N-aryl-, N- aralkyland N-heterocyclic derivatives thereof.

Among the compounds of our invention are included:

17 a-hydroxy-20-oxo-4-pregneno- [3 ,2-c] pyrazole,

6u-chloro-17a-hydroxy-20-0Xo-4-pregneno- [3,2-c] pyrazole,

oz-fluoro-17u-hydroxy-20-oxo-4-pregneno- [3,2-c1pyrazole,

17 ct-hydroxy-6u-methyl-20-oxo-4-pregneno- 3,2-0] pyrazole,

17a-hydroxy-16a-methyl-20-oxo-4-pregneno-[3,2-0] pyr- .azole,

1 7a-hydroxy-16fl-methyl-20-oxo-4-pregneno- 3 ,2-c] pyrazole,

17u-hydroxy-16-methylene-20-oxo-4-pregneno-[3,2c]-

pyrazole,

17 a-acetoxy-ZO-oxo-4-pregneno- 3,2-0] pyrazole,

17 u-acetoxy-6a-chloro-20-oxo-4-pregneno- [3 ,2-c] pyrazole,

17 a-acet0xy-6a-fluoro-20-oxo-4-pregneno- 3,2-c] pyrazole,

17a-acetoxy-6a-methyl-20-oxo-4-pregneno- [3 ,Z-c] pyrazole,

17ot-acetoxy-16a-methyl-20-oxo-4-pregneno- [3 ,2-c]pyrazole,

17ct-hyd roxy-20-oxo-2-phenyl-4-pregneno- [3,2-c] -pyraz ole,

6a-chloro-17a-hydroxy-20-oxo-2-phenyl-4-pregneno [3,2-c1pyrazole,

17 a-hydr oxy-l 6 :x-methyI-Z O-oxo-2'-pl1eny1-4-pre gneno- [3 ,2-c] pyrazole,

17 a-hydroxy- 1 6j3-methyl-20-oxo-2-phenyl-4-pregneno- [3,2-c1pyrazole,

17u-hydroxy-16-rnethylene-20-oxo-2'-phenyl-4-pregneno [3,2-c1pyrazole,

17 e-acetoxy-20-oxo-2'-phenyl-4-pregneno- [3 ,2-c] pyrazole,

17 a-acetoxy-6a-chloro-20-oxo-2'-pheny1-4-pregneno- [3,2-c] pyrazole,

17 a-acetoxy-6a-methyl-20-oxo-2'-phenyl-4-pregneno- [3,2-c] pyrazole,

17a-acet0xy-16a-methyl-ZO-oxo-2'-phenyl-4-pregneno- [3,2-c] pyrazole,

l7a-acetoxy-l 6fi-methyl-20-oxo-2'-phenyl-4-pregneno- [3,2-c1pyrazole,

17 oc-acetoxy-l 6-methylene-20' oxo-Z-phenyl-4-pregneno- [3,2-c] pyrazole,

1 7a-acetoxy-6wchloro-16a-methy1-20-oxo-2'-pl1eny1-4- pregneno- 3,2-c] pyrazole,

17 oz-ac6tOXY-6a-ChlOrO-l 6,8-11ethy1-20- oxo-2'-phenyl-4- pregneno- [3,2-c] pyrazole,

pregneno- 3,2-c] pyrazole,

17a-acetoxy-6a-fluor0-16a-methyl-20-oxo-2'-phenyl-4- pregneno- 3 ,2-c] pyrazole,

17a-acetoxy-6 a-fluoro-l G-methylene -2 O-oxo-2-pheny1-4- pregneno- 3,2-c] pyrazole,

17a-acetoxy-6a, 16a-dimethyl-20-oxo-2'-phenyl-4-pregneno- 3,2-c] pyrazole,

z-3C61OXY-6oe, 1 6/3-dimethyl-ZO-oxo-2'-pheny1-4-preg neno- [3,2-c1pyrazole,

17 a-acetoxy-6oc-methyl- 1 6-me thylene-20-oxo-2'-phenyl-4- pregneno-[3,2-c1pyrazole,

17ot-acetoxy-21-flu0ro-6a,16a-dimethyl-20-oxo-2'-phenyl- 4-pregneno- [3,2-c] pyrazole,

35 17a-acetoxy-6-chloro-16a-methyI-ZO-oxo-2'-phenyl-4,6-

pregnadieno-[3,2-c1pyrazole, 17a-acetoxy-6-chloro- 1 6,B-metl1y1-20-oXo-2'-phenyl- 4,6-

pregnadieno- 17u-hydroxy-20-oX0-2'-phenyl-4,6-pregnadieno- 3,2-c1pyrazole, 17u-acetoXy-6-chlorol 6-methylene-20 -oxo-2'-phenyl-4,6-

pregnadieno- [3 ,Z-c] pyrazole, 17a-acetoxy-6-fiuoro- 16 u-methyl-20-oxo-2'-phenyl-4,6-

pregnadieno-[3,2-c1pyrazole,

pregnadieno- 3,2-c] pyrazole,

17a-acetoXy-6-fiuoro-16-methylene-20-oxo-2-phenyl-4,6-

pregnadieno- 3,2-c] pyrazole,

17a-acetoxy-6,16a-dimethyI-ZO-oxo-2-phenyl-4,6-pregnadieno- [3,2 -c] pyrazole,

17a-acetoxy-6 165-dimethy1-20 oxo-2'-phenyl-4,6-pregnadieno- [3,2-c] pyrazole,

17 a-acetoXy-6-methyl- 1 6-methylene-ZO-oxo-2-phenyl-4,6

pregnadieno-[3,2-c1pyrazole,

17a-acetoXy-6,21-difluoro-20-oXo-2'-phenyl-4,6-pregnadieno- 3,2-c]pyrazole,

17a-acetoxy-21-fiuoro-6,16a-dimethy1-20-oxo-2'-phenyl- 4,6-pregnadieno- [3 ,2-c] pyrazole,

17 a-hydroxy-ZO-oxo-T- p-fluorophenyl) -4,6-pregn adieno- [3 ,2-c] pyrazole,

6-chloro- 17 a-hydroXy-ZO-OXo-Z- (p-fluorophenyl) -4,6-

pregnadieno-[3,2-c1pyrazole,

6-fluoro-17a-hydroxy-20-oxo-2- (p-fiuorophenyl -4,6-

pregnadieno- 3 ,2-c] pyrazole,

17a-hydroxy-6-methyl-20-oxo-2- (p-fluorophenyl) -4,6-

pregnadieno- 3,2-c] pyrazole,

17 a-hydroxy- 1 60t-mthy1-200XO-2'- (p-fiuorophenyl) -4,6-

pregnadieno- [3,2-c] pyrazole,

17 a-hYdIOXY-l 6 ,B-methyl-ZO-oxo-T- p-fiuorophenyl -,4, 6-

' pregnadieno- 3,2-c] pyrazole,

1 7a-hydroXy l 6-methylene-20-oxo-T- (p-fluorophenyl) -4,

6-pregnadieno-[3,2-c]pyrazole, I

17a-acetoxy-20-oxo-Z-(p-fluorophenyl) -4,6-pregnadieno- [3,2-c1pyraz0le,

17a acetoxy-6-chloro-20-oXo-2-(pfiuorophenyl)-4,6-

pregnadieno- [3 ,2 -c] pyrazole,

17a-acetoXy-6-fluoro-20-oXo-2'- (p-fiuorophenyl) -4,6-pregnadieno- [3 ,Z-c] pyrazole, V v i 1 7a-acetoxy- 6-methyl-20-o xo2- (p-fluorophenyl) -4,6-

pregnadieno-[3,2-c1pyrazole,

17a-acetoxy-1 Ga-methyl-ZO-oxo-Z- (p-fluorophenyl) -4,6-

pregnadieno- [3,2-c1pyrazole,

17 a-aceto xy l Gfi-rnethyl-ZO-oxo-Z- (p-fluorophenyl) -4,6-

pregnadieno- [3,2-c1pyrazole, I

17a'-acetoXy-16-methylene-20-oxo-2'-(p-fiuorophenyl) 4,6-pregnadieno- 3 ,Z-c] pyrazole,

l7u acetoxy-6-chloro-1 6u-methyl-20-oxo-2'-(p-fluorophenyl) -4, 6-pregnadieno- [3,2-c1pyrazole,

17a-acetoXy-6-chloro-16B-methyl-20-oXoeZ-(p-fluorophenyl) -4,6-pregnadieno- 3,2-c] pyrazole,

l7a-acetoxy-6-chloro-16-methylene-20-oxo-2- (p-fluorophenyl) -4,6-pregnadien o- [3 ,2-c1pyrazole,

l7a-acetoxy-6-fluoro-16,6-methyl-2 -oxo-2- (p-fluorophenyl) -4,6-pregnadieno- [3,2-c] pyrazole,

17 u-acetoxy-6-fluoro-1 6-methylene-20-oxo-Z- (p-fluorophenyl) -4,6-pregnadieno-[3,2-c1pyrazole,

17 a-acetoxy-6, 1 6a-dimethyl-20-oxo-2'- (p-fluorophenyl) 4,6-pregnadieno- [3 ,2-c] pyrazole,

17a-acetoxyr6-methyl-16-methylene-20-oxo-Z-(p-fluoro- V phenyl) -4,6-pregnadieno- [3,2-c] pyrazole,

17a-acetoxy-21-fiuoro-6,16a-dimethyl-20-oXo-2'-(p-fluorophenyl) -4,6-pregnadieno- [3,2-c1pyrazole,

17a-caproyloXy-20-oxo-4,6-pregnadieno- 3,2-c] pyrazole,

36 17a-caproyloxy-6-chloro-20-oxo-4,6-pregnadieno-[3,2-c]- pyrazole, 17a-caproyloxy-6-iluoro-20-oxo-4,6-pregnadieno-[3,2-01- pyrazole, 17u-caproyloxy-6-methyl-20-oxo-4,6-pregnadieno-[3,2-c1- py 17a caproyloxy-6,16a-dimethyl-4,6-pregnaclieno-[3,2-c]- pyrazole, 17a-acetoxy-20-oXo-19-nor-4,6-pregnadieno-[3,2-c1pyrazole, 17m-acetoXy-6-chloro-20 oxo-19-nor-4,6-pregnadieno- [3,2-c]pyraz ole, I 17e-acetoxy-6-fiuoro-20-oxo-l9-nor-4,6-pregnadieno- [3,2-c]pyrazole, V 17wacetoxy-G-methyl-ZO-oXo-l9-nor-4,6-pregnadieno- [3,2-c1pyrazole, 17a-acetoXy-6,16a-dimethyl-2O-oxo-l9-nor-4,6-pregnadieno- 3,2-c] pyrazole, 6-chloro-17oc-methoxy-20-oxo-4,6-pregnadieno- [3,2-c]- [3,2-c1pyrazole, 17e-methoxy-20-oxo-4,6-pregnadieno-[3,2-c]pyrazole, 6-iluoro-17a-methoXy-20-oXo-4,fi-pregnadieno-[3,2-c1- pyrazole, 17u-methoxy-6-methyl-20-oxo-4,6-pregnadieno-[3,2-0]- pyrazole, 6,16m-dimethyl-17a-methoxy-20-oXo-4,6-pregnadieno- [3,2-c]pyrazole, 17a-methoxy-6,16a-dimethyl-20-oXo-2-pheny1-4,6-preg nadieno-[3,2-c]pyrazole.

Example 28 A spore suspension is prepared in water from a slant of. Aspergillus ochraceus (NRRL 405 and used to inoculate a 250 ml. Erlenmeyer flask containing 50 m1. of medium of the following composition:

7 Percent Edamin 2 Glucose 2 Corn stee'p liquor 0.5

The inoculated flask is incubated at 28 C. on a rotary shaker for 24 hours. The steroid substrate, 170:- acetoxy-ZO-oxo-4-pregneno-[3;2-c]pyrazo1e, is added as a dimethyl-formamide solution mg./ml.) to yield the equivalent of 1.0 g./l. of broth (0.5 ml./flash). After an additional incubation of, 24 hours under the same environmental conditions as the growth phase, the broth is harvested and the desired product isolated.

Fifty milliliters of broth charged with 50 mg. of 17ashows the presence of 17u-acetoXy-1lfi-hydroxy-ZO-oxo- 4-pregneno-[3,2-c]pyrazole.

The reaction product is dissolved in 20 ml. of benzene and charged to a column containing 50 g. of acid washed alumina. The column is washed with benzene and the unreacted starting material eluted with ethyl-benzene mixtures, toobtain 17a-acetoxy-11 3-hydroxy-20-oxo-4-pregneno-[3,2-c]pyrazole.

Various changes and modifications may be made in carrying out the present invention Without departing from the spirit and scope thereof. Insofar as these changes and modifications are within the purview of the annexed claims, they are to be considered as part of our invention.

37 We claim: 1. A compound selected from the group of compounds having structural formulas A and B and the M -analogues of said compounds CHzRi :---ORo pyridyloxide, thiophene and pyrimidyl, and R is selected from the group consisting of lower hydrocarbon carboxylic acyl and lower alkyl.

2. 17a-acetoxy-20-oxo-4-pregneno-[3,2-c]pyrazole.

3. 17a acetoxy 20 0X0 19 nor 4 pregneno- [3,2-c]pyrazole.

4. 17a acetoxy 6a methyl 20 OX0 2 phenyl- 4-pregneno- [3 ,2-c] pyrazole.

5. 17a acetoxy 6,160; dimethyl 20 oxo 4,6- pregnadieno-[3,2-c]pyrazole.

6. 17a methoxy 6,16 dimethyl 20 oxo 2'- phenyl-4,6-pregnadieno-[3,2-c]pyrazole.

7. 17oz acetoxy 4,6 pregnadieno [3,2 c] pyrazole.

8. 17a acetoxy 6 chloro 16a methyl 20 0710- 4,6 pregnadieno [3,2 c]pyrazole.

9. 17oz acetoxy- 6 chloro 16a methyl 2O oxo- 2'-phenyl-4,6-pregnadieno-[3,2-c]pyrazole.

10. acetoxy 6,160; dimethyl 20 oxo 2- phenyl-4,6-pregnadieno- 3 ,2-c] pyrazole.

11. 17a acetoxy 6,165 dimethyl 20 0X0 2'- phenyl-4,6-pregnadieno- [3,2-c] pyrazole.

12. 17a acetoxy 6,165 dimethyl 20 oxo 4,6- pregnadieno- [3,2-c] pyrazole.

13. 17a caproyloxy 6,16oc dimethyl 20 0x0 4,6- pregnadieno-[3,2-c]pyrazole.

14. 17a acetoxy 6 methyl 20 oxo 2 phenyl- 4,6-pregnadieno-[3,2-c]pyrazole.

15. 174x acetoxy 6 chloro 20 OX0 2' phenyl- 4,6-pregnadieno-[3,2-c]py azole.

16. 17a acetoxy 6 chloro 20 0x0 4,6 pregnadieno-[3,2-c]pyrazole.

References Cited in the file of this patent UNITED STATES PATENTS 2,945,852 Bergstrom July 19, 1960 

1. A COMPOUND SELECTED FROM THE GROUP OF COMPOUNDS HAVING STRUCTURAL FORMULAS A AND B AND THE $4,6-ANALOGUES OF SAID COMPOUNDS 